IMMUNOLOGY

MECHANISMS OF IMMUNE
PROTECTION AND
PATHOGENESIS

Dr. Y.Shanti Prabha

 Preface
Immunology stands at the crossroads of modern biology and medicine, providing critical
insights into the body's defense mechanisms and the underlying causes of a wide range of
diseases. This work has been carefully compiled to offer a comprehensive introduction to the
fundamental principles of immunology, while also illuminating its practical implications in
health and disease. Designed to support learners and educators alike, this material progresses
methodically from foundational concepts to more complex immunological processes and clinical
applications.
The structure of this text mirrors the natural progression of immune system understanding
beginning with an overview of immune components and their basic functions, then advancing
through antigen recognition, antibody architecture and utility, immune regulation, and ultimately,
the immune system’s role in pathology and therapeutic intervention. Each unit not only conveys
theoretical knowledge but also integrates applied concepts such as monoclonal antibody
production, hypersensitivity reactions, and immunization strategies, which are vital to current
biomedical research and clinical practice.
This compilation is intended to serve as a foundational scaffold for students of Zoology
and life sciences disciplines, enabling them to grasp core immunological ideas with clarity and
confidence. Through clear exposition and logical organization, it invites learners to appreciate
both the elegance and complexity of immune function and to apply this understanding in both
academic and applied settings.
It is of utmost hope that this compilation not only supports academic success but also
inspires a deeper curiosity and respect for the remarkable system that defends the human body
with such precision and adaptability.
 Contents

1. Introduction to basic concepts in Immunology

2. Cells of immune system
3. Organs of immune system
4. Innate and adaptive immunity
5. Basic properties of antigens and Introduction to types of antigens
6. B and T cell epitopes, paratopes
7. Haptens and adjuvants
8. Factors influencing immunogenicity
9. Structure of antibody
10. Classes of antibodies and Functions of antibodies
11. Hybridoma technology and Concept of Monoclonal antibodies.
12. Structure and functions of Major Histocompatibility Complexes
13. Basic properties and functions of the cytokines.
14. Exogenous and Endogenous pathway of antigen presentation and processing
15. Gell and Coombs’ classification and various types of hypersensitivities
16. Introduction to concepts of autoimmunity, Primary and secondary immunodeficiencies.
17. General introduction to vaccines, Types of vaccines, Immunization programme
18. Organ transplantation- Graft rejection, immune suppressors
 Introduction to basic concepts in Immunology

Immunology is the study of the immune system, which plays a crucial role in protecting
the body from infections, diseases, and harmful substances. The immune system is a highly
complex network of cells, tissues, organs, and molecules that work together to defend the body
from pathogens (microorganisms like bacteria, viruses, fungi, and parasites) while maintaining
tolerance to the body’s own cells. The immune system consists of two primary components
namely the innate immune system and the adaptive immune system. The adaptive immune
system is further subdivided into two branches called as humoral immunity, mediated by B
cells and antibodies, and cell-mediated immunity, driven by T cells. These systems work
collaboratively to protect the body from invaders.
A significant feature of the immune system is its ability to differentiate between the
body’s own cells and foreign substances. During early development, immune cells that bind to
self-antigens are eliminated to prevent autoimmunity. On the other hand, immune cells that
recognize and bind to foreign antigens are activated, undergo clonal selection, and proliferate to
mount a defense against the pathogen.

History of Immunology:
The origins of immunology trace back thousands of years, where foundational practices
and observations began shaping this essential scientific field. Ancient cultures in regions like
China and India developed early forms of vaccination by introducing small amounts of infectious
material into individuals to safeguard against illnesses such as smallpox. In the late 1700s,
Edward Jenner revolutionized immunology by showing that cowpox inoculation could
successfully prevent smallpox, marking the advent of modern vaccination techniques. Building
upon Jenner's work, Louis Pasteur made significant strides in the 19th century by creating
vaccines for diseases like rabies and anthrax. He also established the germ theory, which
emphasized the role of microorganisms in infectious diseases. The discovery of antibodies by
Emil von Behring and Shibasaburo Kitasato further illuminated the concept of humoral
immunity, while Ilya Metchnikoff's study of phagocytosis offered insights into cellular
immunity. The 20th century saw monumental progress in immunology, including the
identification of the major histocompatibility complex (MHC), the introduction of the clonal
selection theory by Frank Macfarlane Burnet, and the emergence of immunosuppressive
treatments that transformed organ transplantation. In contemporary times, immunology continues
to advance through innovations like monoclonal antibodies, gene editing, and immunotherapy,
addressing a wide spectrum of diseases, including infectious illnesses and cancer. This evolution
showcases the remarkable progression of immunology from ancient practices to an advanced,
specialized scientific discipline central to modern healthcare and medicine.

Chronological sequence of key discoveries, advancements, and milestones in

the field of immunology:
 Ancient Practices (Circa 2000 BCE): Early forms of immunization were observed in

ancient civilizations such as China and India. Methods like variolation involved the
intentional exposure to small amounts of infectious material to provide protection against
diseases such as smallpox.
 1796 – Edward Jenner: Edward Jenner developed the first modern vaccine by

demonstrating that cowpox inoculation could prevent smallpox, marking the birth of
vaccination and modern immunology.
 1860s – Louis Pasteur: Louis Pasteur established the germ theory of disease, proving

microorganisms cause infections. He also developed vaccines for rabies and anthrax,
building the foundation for microbiological immunology.
 1890 – Emil von Behring and Shibasaburo Kitasato: Discovered antibodies,

introducing the concept of humoral immunity, which involves the production of specific
molecules to neutralize pathogens.
 1891 – Ilya Metchnikoff: Metchnikoff identified phagocytosis, demonstrating how cells

like macrophages engulf and destroy pathogens, forming the basis of cellular immunity.
 1930s – Discovery of Complement System: Advances in understanding the complement

system, which enhances immune responses, were made during this era.
 1958 – Clonal Selection Theory: Frank Macfarlane Burnet proposed the clonal

selection theory, explaining how immune cells are selected to respond to specific
antigens and proliferate upon activation.
  1960s – Major Histocompatibility Complex (MHC): MHC molecules were identified,

revealing their essential role in antigen presentation and immune system regulation.
 1970s – Monoclonal Antibodies: Georges Köhler and César Milstein pioneered

monoclonal antibody technology, enabling the production of specific antibodies for

therapeutic and research purposes.
 1980s – HIV/AIDS: Immunology gained greater attention due to the emergence of

HIV/AIDS, highlighting the consequences of immune system failure.

 1990s – Immunotherapy: Innovations in cancer treatment led to the development of

immunotherapies, such as monoclonal antibodies and checkpoint inhibitors.

 21st Century – Gene Editing and mRNA Technology: Recent breakthroughs include

the application of CRISPR-Cas9 for manipulating immune system components and the
advent of mRNA vaccines, which proved pivotal in combating COVID-19.

Key concepts in Immunology:

1. Innate and Adaptive Immunity - The immune system is broadly classified into innate and
adaptive immunity, which work together to protect the body from harmful invaders. Innate
immunity is the first line of defense and is non-specific, providing immediate protection
against a wide range of pathogens. It includes physical barriers like skin and mucous
membranes, chemical barriers like acidic gastric secretions and enzymes, and cellular
components like macrophages and natural killer (NK) cells. Innate immunity does not have
memory, meaning it reacts the same way regardless of previous encounters with pathogens.
Adaptive immunity, on the other hand, is highly specific and develops a memory. It
becomes activated when the innate immune system is overwhelmed. Adaptive immunity
involves B cells that produce antibodies to neutralize antigens (humoral immunity) and T
cells that directly attack infected cells or coordinate immune responses (cell-mediated
immunity). This memory ensures a faster and stronger response upon subsequent exposure to
the same pathogen.
2. Self vs. Non-Self Recognition - One of the immune system's most vital features is its ability
to distinguish between self and non-self. During early development, immune cells undergo a
rigorous process to learn this distinction. Cells that react to self-antigens are eliminated
through a process known as negative selection, preventing autoimmune diseases where the
 immune system attacks the body's own tissues. Only immune cells that recognize foreign
antigens are allowed to mature and function, ensuring efficient defense against invaders. This
ability to discriminate is mediated by molecules encoded by the Major Histocompatibility
Complex (MHC). MHC Class I molecules present antigens to cytotoxic T cells (CD8+),
while MHC Class II molecules present antigens to helper T cells (CD4+). The interaction of
MHC molecules with immune cells is critical for antigen recognition and immune response.
3. Humoral and Cell-Mediated Immunity - Adaptive immunity is divided into humoral
immunity and cell-mediated immunity, each playing unique roles. Humoral immunity is
mediated by B cells, which produce antibodies that circulate in the blood and neutralize
pathogens by binding to their antigens. Antibodies also facilitate opsonization, where
pathogens are marked for destruction by phagocytic cells. Cell-mediated immunity involves
T cells that attack infected or abnormal cells. Helper T cells (CD4+) release cytokines to
activate other immune cells, while cytotoxic T cells (CD8+) directly kill infected or
cancerous cells by inducing apoptosis. Together, these two branches of adaptive immunity
ensure a comprehensive defense against diverse types of threats.
4. The Complement System is an integral part of innate immunity consisting of proteins that
enhance immune responses. Complement proteins can directly kill pathogens by forming a
membrane attack complex (MAC), which creates pores in the pathogen's membrane.
Complement also aids in opsonization, marking pathogens for destruction by phagocytes, and
amplifies inflammation through the release of immune mediators. These processes
collectively ensure a rapid and effective response to infections.
5. Antigen Presentation is a process through which specialized cells known as antigen-
presenting cells (APCs) display pathogen-derived antigens to T cells. This is essential for
activating adaptive immunity. APCs, including dendritic cells, macrophages, and B cells,
present antigens using MHC molecules, ensuring that T cells recognize and respond to the
pathogen. This process is central to coordinating immune responses and establishing long-
term immunity.
6. Immunological Memory is a key feature of the adaptive immune system that distinguishes it
from innate immunity. After an initial encounter with a pathogen, memory B and T cells are
generated. These cells retain the information needed to recognize the pathogen and mount a
rapid and robust immune response during subsequent exposures. This concept forms the
 basis of vaccination, where exposure to a harmless form of the pathogen stimulates the
formation of memory cells, protecting the body from future infections. The primary immune
response is slower and weaker compared to the secondary immune response, which is faster
and more efficient due to the presence of memory cells.
7. Hypersensitivity - Hypersensitivity refers to exaggerated or inappropriate immune responses
that can damage the body's own tissues. It is categorized into four types: Type I
(Immediate): Mediated by IgE antibodies, causing allergic reactions such as asthma or
anaphylaxis.Type II (Cytotoxic): Involves IgG or IgM antibodies targeting self cells, leading
to cell destruction. Type III (Immune Complex): Occurs when antigen-antibody complexes
deposit in tissues, triggering inflammation and tissue damage (e.g., lupus). Type IV
(Delayed): Mediated by T cells, causing inflammation such as in contact dermatitis.
8. Autoimmune Diseases arise when the immune system fails to recognize self and attacks the
body's own tissues. Conditions like rheumatoid arthritis, lupus, and type 1 diabetes result
from this breakdown in self-tolerance. The causes are multifactorial, involving genetic
predispositions, environmental triggers, and immune dysregulation. Understanding the
mechanisms underlying autoimmune diseases has led to therapies aimed at suppressing or
modulating immune activity, offering hope to patients.
9. Vaccination is one of the greatest achievements in immunology, leveraging the concept of
immunological memory. Vaccines expose the immune system to harmless antigens derived
from pathogens, stimulating the production of antibodies and memory cells without causing
disease. This prepares the immune system to respond effectively to future encounters with
the real pathogen. Vaccines can be live attenuated (weakened forms of the pathogen),
inactivated (killed pathogens), subunit-based, or mRNA-based. They have been instrumental
in controlling diseases like polio, measles, and COVID-19.
 Cells of the immune system

The immune system consists of various organs and tissues spread throughout the body. These
immune organs are divided into two main categories:
A. Primary lymphoid organs: These are where lymphocytes (a type of white blood cell) develop
and mature.
B. Secondary lymphoid organs: These trap antigens (foreign substances) from the tissues and
blood, and are where lymphocytes actually interact with antigens.
All these organs are connected by the lymphatic system and blood vessels, forming a functional
unit. The immune system also includes different types of white blood cells, such as lymphocytes (T-
cells, B-cells, and natural killer cells), neutrophils, monocytes, and macrophages. These cells play a key
role in defending the body against foreign elements.
Of all these cell types, only lymphocytes exhibit diversity, specificity, memory, and the ability
to distinguish self from non-self, which are key features of adaptive immune responses. The other white
blood cells support adaptive immunity by activating lymphocytes, enhancing the removal of antigens
through phagocytosis, or secreting immune-effector molecules. Certain white blood cells release
cytokines, which regulate immune responses. Other crucial proteins in the immune system are
antibodies produced by B-lymphocytes and complement proteins, which are activated by antibodies.

Cells of the Immune System

Image source: Riott’s Essential Immunology
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Types of Immune System Cells:

The immune system's central cells, lymphocytes, are in charge of adaptive immunity as well as
the immunological traits of self-and non-self recognition, diversity, specificity, and memory. The other
immune cells' roles include presenting antigens, secreting cytokines, and engulfing and eliminating
microorganisms.
Lymphocytes:
Lymphocytes are a type of white blood cell, making up 20-40% of all white blood cells in the
body and 99% of the cells in the lymph. There are about 100 billion lymphocytes in the human body.
They constantly move through the blood and lymph, allowing them to migrate into body tissues and
lymphoid organs, thereby playing a key role in connecting and integrating the immune system.
Lymphocytes are classified into three main types based on their functions and cell-membrane
components:

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1. B-lymphocytes (B-cells):
★ Referred to as CD19 or CD20 cells in lab reports.
★ Their primary role is to produce antibodies, also known as immunoglobulins.
★ B-lymphocytes develop and mature in the bone marrow from hematopoietic stem cells.
★ After maturation, they form unique antigen-binding receptors called B-cell receptors (or
antibodies) on their membranes.
★ Mature B-cells migrate to the bone marrow, lymph nodes, spleen, intestines, and
bloodstream.
★ When a naive B-cell encounters an antigen for the first time, it binds to the antigen using
its antibodies, causing the B-cell to rapidly divide and differentiate into memory B-cells
and plasma cells.
★ Memory B-cells have a longer lifespan and express the same antibodies as the parent B-
cells.
★ Plasma cells produce and secrete large amounts of antibodies into the bloodstream,
tissues, and various secretions.
★ Antibodies are specialized proteins designed to target specific antigens like a lock and
key.
★ Each plasma cell produces only one type of antibody.
★ During maturation, B-cells learn not to produce antibodies against healthy tissues.
★ Antibodies vary in function and chemical structure, which determines their class.

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T-Lymphocytes :
T-lymphocytes are also known as T-cells, often named in lab reports as CD3 cells They also
arise in the bone marrow but migrate to the thymus gland for maturation, where they express a unique
antigen-binding molecule on its membrane known as the T-cell receptor. The name T originated from
its site of maturation, the Thymus. Mature T-cells leave the thymus and populate other organs of the
immune system, such as the spleen, lymph nodes, bone marrow, and blood. Unlike the B-cell receptors
that can recognize antigens alone, T-cell receptors only recognize antigens that are bound to cell
membrane proteins known as Major Histocompatibility Complex (MHC) molecules. The MHC
molecule recognizes antigens that are presented to them by antigen-processing cells (APCs) on their
cell membrane. The two major classes of MHC molecules are Class I MHC molecules, which are
expressed by nearly all nucleated cells of vertebrate species, consist of a heavy chain linked to a small
invariant protein called 2-microglobulin. Class II MHC molecules, which consist of an alpha and a beta
glycoprotein chain, are expressed only by antigen-presenting cells. When a naive T cell encounters an
antigen combined with an MHC molecule on a cell, the T cell proliferates and differentiates into
memory T cells and various effector T cells.

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 T-lymphocytes, also known as T-cells (CD3 cells in lab reports), originate in the bone marrow
and migrate to the thymus for maturation. In the thymus, they develop a unique antigen-binding
molecule on their membrane called the T-cell receptor. The name "T-cell" comes from their thymus
maturation site. Mature T-cells leave the thymus and spread to other immune system organs, such as
the spleen, lymph nodes, bone marrow, and blood. Unlike B-cell receptors that can recognize antigens
directly, T-cell receptors only recognize antigens bound to Major Histocompatibility Complex (MHC)
molecules on cell membranes. There are two major classes of MHC molecules:
★ Class I MHC molecules
★ Class II MHC molecules

MHC Class I proteins are expressed on most nucleated cells and present antigens from
intracellular pathogens to cytotoxic T cells for cell killing. On the other hand, MHC Class II proteins
are restricted to antigen-presenting cells and present antigens from extracellular pathogens to helper
T cells for immune regulation and activation.

MHC Class I and MHC Class II Molecules

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When a naive T-cell encounters an antigen presented with an MHC molecule, it proliferates and
differentiates into memory T-cells and various effector T-cells. T-cells are divided into three types: T
helper (Th), T cytotoxic (Tc), and T suppressor (Ts) cells. Th and Tc cells are distinguished by the
presence of CD4 and CD8 membrane glycoproteins, respectively. T cells with CD4 are Th cells, while
those with CD8 are Tc cells. Th cells recognize antigens presented by MHC class II molecules and,
upon activation, secrete growth factors called cytokines. These cytokines play a crucial role in
activating B-cells, Tc cells, macrophages, and other immune cells. The specific cytokine patterns
produced by activated Th cells lead to different immune responses. Th-derived cytokines help Tc cells
recognize antigens presented by MHC class I molecules, leading to the proliferation and differentiation
of Tc cells into Cytotoxic T-lymphocytes (CTLs). Tc cells, unlike helper T-Cells, do not induce
cytokine secretion but exhibit cell-killing activity. CTLs are essential in monitoring and eliminating
cells displaying antigens, such as tumor cells, virus-infected cells, and foreign tissue graft cells. CTLs
target altered self-cells presented by MHC class I molecules.

NK cells, or natural killer cells:

Unlike the B and T-cell lineages, these large granular lymphocytes do not exhibit surface
markers. They were initially identified in 1976 based on signs of a limited population of large granular
lymphocytes that, in the absence of prior tumour immunisation, exhibited a cytotoxic effect against a
variety of tumour cells. The host's defence against tumour cells and cells infected with some, but not
all, viruses is bolstered by these cells. They make about 5–10% of the peripheral blood lymphocytes in
humans.
Natural Killer (NK) cells recognize antigens through two main mechanisms:
1. NK cell receptors detect abnormalities like reduced expression of class I MHC molecules and
unusual surface antigens on tumor cells and virus-infected cells.
2. NK cells also identify tumor cells and virus-infected cells by recognizing antigens against
which the immune system has already produced antibodies. These antibodies, binding to the
target cell surfaces, are recognized by NK cells' CD16 receptors (which bind to the Fc region of
IgG antibodies). The NK CD16 receptors attach to these antibodies and destroy the target cells
through Antibody-dependent cell-mediated cytotoxicity (ADCC).
NK cells play a critical role in defending against tumors. For instance, in humans, Chediak-
Higashi syndrome, an autosomal recessive disorder, is linked to defects in neutrophils, macrophages,
and NK cells, resulting in a higher incidence of lymphomas. Similarly, mice with the autosomal
mutation called beige lack NK cells and are more prone to tumor growth after injection with live tumor
cells.
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natural-killer-NK-cell-response-according-to-the-missing-self-and.png

There are special NK cells known as NK1-T cells, which share features of both T-lymphocytes
and NK cells. These cells have T-cell receptors (TCRs) that interact with CD1 molecules, unlike
normal TCRs that interact with class I or II MHC molecules. Like NK cells, they have CD16 and other
NK receptors that enable them to kill cells.
NK1-T cells rapidly secrete large amounts of cytokines, supporting antibody production by B-
cells, inflammation, and the development and expansion of cytotoxic T-cells. Some immunologists see
these cells as a rapid response system providing early help while conventional T helper (Th) responses
develop.
Mononuclear phagocytes:
Mononuclear phagocytes include immune cells such as monocytes, which circulate in the blood,
and macrophages, which are found in tissues. During hematopoiesis in the bone marrow, granulocyte-
monocyte progenitor cells differentiate into promonocytes. These promonocytes then leave the bone
marrow, enter the blood, and further differentiate into mature monocytes.
Monocytes circulate in the bloodstream for about 8 hours, during which they enlarge. They then
migrate into tissues and differentiate into specific tissue macrophages or dendritic cells. This
differentiation involves several changes:
 The cell enlarges five to tenfold.
 Its intracellular organelles increase in number and complexity.
  It gains increased phagocytic ability and produces higher levels of hydrolytic enzymes.
 It begins to secrete various soluble factors.

Structure of Monocyte
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Macrophages

Macrophages are spread throughout the body. Some become fixed macrophages residing in
specific tissues, while others remain motile, called free or wandering macrophages. Free macrophages
move throughout the tissues using amoeboid movement. Macrophage-like cells perform various
functions in different tissues and are named based on their location:
1) Alveolar macrophages in the lungs.
2) Histiocytes in connective tissues.
3) Kupffer cells in the liver.
4) Mesangial cells in the kidneys.
5) Microglial cells in the brain.
6) Osteoclasts in bones.

Structure of Macrophage
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Macrophages are usually in a resting phase but can be activated by several immune responses.
Initial activation often occurs through the phagocytosis of certain antigens. However, macrophage
activity can be further enhanced by cytokines secreted by activated T helper (TH) cells, inflammatory
response mediators, and bacterial cell wall components. Interferon-gamma, secreted by activated TH
cells, is one of the most potent macrophage activators.
Activated macrophages are more effective at eliminating potential pathogens than resting
macrophages. They exhibit greater phagocytic activity, an increased ability to kill ingested microbes,
increased secretion of inflammatory mediators, and an enhanced ability to activate T cells.
Additionally, activated macrophages secrete various cytotoxic proteins that help eliminate a broad
range of pathogens, including virus-infected cells, tumor cells, and intracellular bacteria. Activated
macrophages also express higher levels of class II MHC molecules, allowing them to function more
effectively as antigen-presenting cells.
Some functions of macrophages include:
✿ Phagocytosis: Engulfing bacteria, viruses, and other foreign particles. Macrophages have Fc
receptors that interact with the Fc component of IgG, facilitating the ingestion of opsonized
organisms. They also have receptors for C3b, another important opsonin. After ingestion, the
phagosome fuses with a lysosome, and the microbe within is killed by reactive oxygen, reactive
nitrogen compounds, and lysosomal enzymes.
✿ Antimicrobial and cytotoxic activities: Includes both oxygen-dependent and oxygen-
independent killing.
✿ Antigen processing: After ingesting and degrading foreign materials, antigen fragments are
presented on the macrophage cell surface with class II MHC proteins for interaction with CD4+
helper T cells. The degradation of the foreign protein stops following the association with class
II MHC proteins in the cytoplasm. The complex is then transported to the cell surface by
transporter proteins.
✿ Secretion of growth factors: Important for the development of an immune response, including
cytokines like interleukin 1 (IL-1), TNF-α, and interleukin 6 (IL-6), which promote
inflammatory responses, complement proteins, hydrolytic enzymes, and a cascade of Tumor
Necrosis Factors (TNF-α) that induce and kill tumor cells and promote hematopoiesis.
 Macrophage function
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Granulocytes
These are a type of white blood cell (leukocytes). They are classified by their cellular shapes
and how their cytoplasm stains. This group includes neutrophils, eosinophils, basophils, and mast cells.
All granulocytes have distinctive multilobed nuclei, making them easy to identify compared to
lymphocytes, which have round nuclei. The cytoplasm of granulocytes contains numerous granules that
release proteins when they encounter pathogens. These proteins have various roles, including directly
damaging pathogens, regulating other white blood cells, and helping to remodel tissues at infection
sites.
1) Neutrophils have a multilobed nucleus and granulated cytoplasm that stains with both acid and
basic dyes, earning them the name polymorphonuclear leukocytes (PMNs).
2) Eosinophils have a bilobed nucleus and granulated cytoplasm that stains red with the acid dye
eosin.
3) Basophils have a lobed nucleus and granulated cytoplasm that stains blue with the basic dye
methylene blue.
 Neutrophils and eosinophils are phagocytic, meaning they can engulf and digest pathogens,
whereas basophils are not. Neutrophils are the most abundant, making up 50-70% of circulating
leukocytes, while eosinophils make up 1-3%, and basophils and mast cells each make up less than 1%.
Neutrophils are produced in the bone marrow through hematopoiesis. Once released into the
bloodstream, they circulate for 7–10 hours before migrating into tissues, where they live for only a few
days. During infections, a large number of neutrophils are produced, and they are typically the first
cells to arrive at the inflammation site. This temporary increase in circulating neutrophils, known as
leukocytosis, is a medical indicator of infection. The process of neutrophils moving from the
bloodstream into tissues is called extravasation, which occurs in several steps:
★ Adherence to the vascular endothelium.
★ Penetration between endothelial cells lining the vessel wall.
★ Penetration through the vascular basement membrane into tissue spaces.

During inflammation, various substances act as chemotactic factors, promoting the

accumulation of neutrophils at the inflammation site. These factors include complement components,
blood-clotting system components, and cytokines secreted by activated T helper cells and macrophages.

Neutrophil
Image source: Image source: Kuby Immunology 7th Edition

Neutrophils function as active phagocytes, similar to macrophages. The phagocytosis

mechanism in neutrophils involves primary and secondary granules containing lytic enzymes and
bactericidal substances. Primary granules, a type of lysosome, contain peroxidase, lysozyme, and
hydrolytic enzymes, while secondary granules contain collagenase, lactoferrin, and lysozyme. Both
granule types fuse with phagosomes to digest and eliminate their contents.
 Neutrophils use both oxygen-dependent and oxygen-independent pathways to produce
antimicrobial substances. They exhibit a larger respiratory burst than macrophages, generating more
reactive oxygen and nitrogen intermediates. Additionally, neutrophils express higher levels of defensins
compared to macrophages.

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Eosinophils are mobile, phagocytic cells that can migrate from the bloodstream into tissue
spaces. While they have a phagocytic role in eliminating antigens, their significance in this function is
less than that of neutrophils. Eosinophils are involved in defending against multicellular parasitic
organisms, such as worms. The contents of their granules can damage parasite membranes, and they
often cluster around invading worms, releasing proteins that harm these parasites.
In addition to their role in parasitic defense, eosinophils, like neutrophils and basophils, can
secrete cytokines that regulate B and T lymphocytes, influencing the adaptive immune response. In
regions where parasitic infections are less common, eosinophils are more recognized for their
contributions to asthma and allergy symptoms.

Basophils are non-phagocytic granulocytes with large granules filled with basophilic proteins
that stain blue with standard H & E staining. They are naturally present in the body's circulation and
can be very potent. Basophils work by binding to circulating antibodies and releasing the contents of
their granules, which are pharmacologically active substances in their cytoplasm. These substances play
a key role in allergic responses. For example, histamines, which are common in basophilic granules,
increase blood vessel permeability and smooth muscle activity. Like eosinophils, basophils are
important in responding to parasites, particularly helminths (worms). Basophils also secrete cytokines
that help modulate the adaptive immune response.

Basophil
Image source: Kuby Immunology 7th Edition

Mast Cells
Mast cells originate in the bone marrow and are released into the bloodstream as
undifferentiated cells. They mature upon entering various tissues, where they can be found in the skin,
connective tissues of various organs, and mucosal epithelial tissues of the respiratory, genitourinary,
and digestive tracts. Like basophils, mast cells contain numerous cytoplasmic granules filled with
histamine and other pharmacologically active substances.
Mast cells play a crucial role in allergy development. Although basophils and mast cells share
many similarities, their exact relationship is not entirely understood. Some theories suggest that
basophils are the blood-borne equivalent of mast cells, while others propose that they have distinct
origins and functions.

Image source: Kuby Immunology 7th Edition

Dendritic cells:
Dendritic Cells were discovered by Ralph Steinman in the mid-1970s, and he was awarded the
Nobel Prize in 2011 for this discovery. These cells are named for their long membrane extensions that
resemble the dendrites of nerve cells. These extensions dynamically extend and retract, increasing the
surface area available for interacting with lymphocytes. Dendritic cells are diverse and originate from
both myeloid and lymphoid lineages of hematopoietic cells.

Dendritic cell
Image source: Kuby Immunology 7th Edition

Dendritic cells perform two key functions: capturing antigens in one location and presenting
them in another. Immature dendritic cells patrol the body for pathogens outside the lymph nodes,
capture foreign antigens, process them, and then migrate to the lymph nodes to present the antigens to
naive T cells, initiating the adaptive immune response. As guards in peripheral tissues, immature
dendritic cells capture antigens through:
1) Phagocytosis
2) Receptor-mediated endocytosis
3) Pinocytosis

During maturation, dendritic cells transition from an antigen-capturing phenotype to one

specialized for antigen presentation. They lose some functions, like phagocytosis and large-scale
pinocytosis, but gain a significantly increased ability to present antigens and express costimulatory
molecules essential for activating naive T cells. Once activated, dendritic cells leave peripheral tissues,
enter the blood or lymphatic circulation, and migrate to lymphoid organs where T cells reside to present
antigens.

There are various types of dendritic cells, but their main function is to present antigens to T
helper (TH) cells. The four types of dendritic cells are:
a) Langerhans cells
b) Interstitial dendritic cells
c) Myeloid cells
d) Lymphoid dendritic cells

Each type originates from hematopoietic stem cells through different pathways and locations.
Despite their differences, they all express high levels of class II MHC molecules and co-stimulatory B7
family members. This makes them more potent antigen-presenting cells than macrophages and B cells,
which need activation to function as APCs.
Immature or precursor forms of these dendritic cells capture antigens via phagocytosis or
endocytosis. Once processed, the mature dendritic cells present the antigens to TH cells. During
microbial invasion or inflammation, both mature and immature Langerhans and interstitial dendritic
cells migrate to lymph nodes, where they present antigens to TH cells, initiating an immune response.
Follicular dendritic cells, which do not originate in the bone marrow, have a different role. They
do not express class II MHC molecules and therefore do not function as APCs for TH-cell activation.
Found exclusively in lymph node follicles, which are rich in B cells, these cells have high levels of
membrane receptors for antibodies. This allows them to bind antigen-antibody complexes, influencing
B cell responses.
 Organs of the immune system
Introduction:
The immune system comprises a complex and essential network of various organs and
tissues, each with different functions in developing immune responses. These organs, known as
lymphoid organs, are involved in the growth, development, and deployment of lymphocytes.
Functionally, they are categorized into two main groups: primary and secondary lymphoid
organs. These organs are interconnected by the body's blood vessels and lymphatic systems,
creating a unified functional unit during immune responses against antigens by transporting
lymphocytes throughout the body, leading to systemic immunity. Additionally, tertiary lymphoid
tissues can import lymphoid cells during inflammatory responses.
1. Primary Lymphoid Organs:
These organs provide the environment for the development and maturation of lymphocytes
(e.g., white blood cells, leukocytes). The thymus and bone marrow are the primary (or central)
lymphoid organs. Immature lymphocytes generated during hematopoiesis mature and commit to
specific antigens in these primary organs. Mature lymphocytes residing in primary lymphoid
organs are immunocompetent, meaning they can mount an immune response.

Lymphoid organs
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Are-Lymphoid-Organs.png
1.1. Bone Marrow:
Bone marrow is the origin and development site for B-cells in most mammals, including
humans and mice. In birds, B-cell maturation occurs in the “bursa of Fabricius.” Mammals like
primates and rodents don't have a similar primary lymphoid organ, but in early gestation in cattle
and sheep, the fetal spleen hosts B-cell maturation, proliferation, and diversification. Later, this
function shifts to the ileal Peyer’s patch in the intestine, which contains a large number of B cells
(over 10 billion). Immature B cells proliferate and differentiate in the bone marrow after arising
from lymphoid progenitors. Stromal cells in the bone marrow interact directly with B cells and
secrete cytokines necessary for further development. During maturation, B cells with self-
reactive antibody receptors are eliminated through a selection process in the bone marrow.

Bone marrow
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1.2. Thymus:
The thymus is the first organ to produce lymphocytes during development, providing an
environment for T cell development and maturation. It's a flat, bilobed organ located above the
heart, with each lobe divided into lobules by connective tissue called trabeculae. Each lobule has
two compartments: the outer cortex, densely packed with immature T cells (thymocytes), and the
inner medulla, which has fewer thymocytes. Thymic lymphocytes are surrounded by epithelial
cells, dendritic cells, and macrophages.
In the outer cortex, some thymic epithelial cells, known as nurse cells, surround
thymocytes to form large complexes that influence their development. Dendritic cells and
macrophages are found in the medulla and at the junction between the cortex and medulla.
Epithelial cells in the thymus produce various cytokines needed for the differentiation of thymic
precursors into mature T cells. Thymocytes are attracted to the thymus from the bone marrow by
these cytokines. They mature in the cortex, migrate to the medulla, and are eventually released to
enter peripheral lymphoid tissues.
About 75% of all lymphocytes in the thymus are located in the deeper cortex.
Thymocytes in this area express CD1, CD4, and CD8 membrane molecules, unlike blood T cells,
which express either CD4 or CD8. Thymocytes undergo thymic selection, where T cells that
recognize self molecules are removed through apoptosis. Consequently, most T cells produced in
the thymus die. As they migrate to the medulla, they lose either CD4 or CD8 expression due to
genetic rearrangement. These naive, mature T cells then enter peripheral blood circulation and
are transported to secondary lymphoid organs, where they encounter and respond to foreign
antigens.

TS of Thymus
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2. Secondary Lymphoid Organs:
These organs capture antigens from specific tissues or vascular spaces and provide sites
for mature lymphocytes to interact with antigens effectively. The lymph nodes and spleen are the
primary secondary (or peripheral) lymphoid organs in the immune system.
2.1. Lymph Nodes:
Lymph nodes are encapsulated, bean-shaped structures that contain a reticular network of
lymphocytes, macrophages, and dendritic cells. Their main function is to filter antigens. Located
at the junctions of major lymphatic vessels, lymph nodes are the first organized structures where
immune responses to tissue antigens are initiated. Lymphatic vessels transport lymph to the
nodes, where antigens are filtered out. As the lymph is filtered in the nodes, it becomes enriched
with antibodies, cytokines, and mainly T lymphocytes.
Morphologically, a lymph node can be divided into three main concentric regions: the
cortex, paracortex, and medulla, each providing a distinct microenvironment.
a) Cortex: The outermost layer, mainly composed of B lymphocytes, macrophages, and
follicular dendritic cells arranged in clusters called primary follicles. Upon antigen
activation, these primary follicles enlarge into secondary follicles with a germinal center,
densely packed with proliferating B lymphocytes and plasma cells, along with
macrophages and dendritic cells. The germinal center is a crucial site for B-cell activation
and differentiation into plasma cells and memory cells.
b) Paracortex: Located beneath the cortex, this region is primarily populated with T

lymphocytes and some interdigitating dendritic cells. These dendritic cells express high
levels of class II MHC molecules, essential for presenting antigens to T helper (TH) cells.
c) Medulla: The innermost layer, sparsely populated with lymphoid-lineage cells, mainly
plasma cells actively secreting antibodies, along with activated TH and TC cells. This
region has a high concentration of immunoglobulins (Ig) due to the abundance of plasma
cells.
 Structure of Lymph node
Image source: Janeway’s Immunology 9th edition

Lymph enters the lymph node through afferent lymphatic vessels, which pierce the node's
capsule at various points and empty into the subcapsular sinus. The lymph then flows slowly
through the cortex, paracortex, and medulla, allowing phagocytic cells and dendritic cells to
capture any bacteria or particulate material (e.g., antigen-antibody complexes) carried by the
lymph. The lymph exits the node through a single efferent lymphatic vessel, which is enriched
with antibodies and has a higher concentration of lymphocytes compared to the afferent vessels.
2.2. Spleen:
The spleen is a large, oval secondary lymphoid organ located high in the left abdominal
cavity. It filters blood and traps blood-borne antigens, responding to systemic infections. Blood-
borne antigens and lymphocytes enter the spleen through the splenic artery. The spleen is
encapsulated and extends projections called trabeculae, creating a compartmentalized structure
with two types of compartments: red pulp and white pulp, separated by a marginal zone.
 Red Pulp: Contains red blood cells (erythrocytes) mixed with macrophages, dendritic

cells, a few lymphocytes, and plasma cells. This is where old and defective red blood
cells are destroyed and removed.
  White Pulp: Surrounds the splenic artery, forming a periarteriolar lymphoid sheath

(PALS) mainly populated by T lymphocytes. The marginal zone, peripheral to the PALS,
is rich in lymphocytes and macrophages organized into primary lymphoid follicles.

The splenic artery carries blood-borne antigens and lymphocytes to the spleen, emptying
them into the marginal zone. Interdigitating dendritic cells trap the antigens and present them
with class II MHC molecules to T helper (TH) cells. The activated TH cells then further activate
B cells. Together with some TH cells, the activated B cells migrate to primary follicles in the
marginal zone and develop into secondary follicles with germinal centers, similar to those in
lymph nodes.

TS of Spleen
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1483.png?ezimgfmt=ng:webp/ngcb1
3. Tertiary Lymphoid Tissues:
Tertiary lymphoid tissues generally contain fewer lymphoid cells than secondary
lymphoid organs. However, they play a crucial role in immune responses by rapidly increasing
lymphoid cell numbers during such responses. Key examples include mucosa-associated
lymphoid tissue (MALT), cutaneous-associated lymphoid tissue (CALT), and intraepithelial
lymphocytes (IEL).
3.1. Mucosal-Associated Lymphoid Tissue (MALT):
Mucous membranes lining the digestive, respiratory, and urogenital systems are
susceptible to pathogen attacks. These linings are protected by clusters of non-encapsulated,
organized lymphoid tissues known as MALT. These tissues range from loosely clustered
lymphoid cells in the lamina propria of intestinal villi to well-organized structures like the
tonsils, appendix, and Peyer's patches of the intestinal submucosa.
The lamina propria is rich in macrophages, B cells, plasma cells, and activated T helper
(TH) cells, providing stronger immunity compared to other body parts. MALT's critical role in
the body's defense is due to its large population of antibody-producing plasma cells. Notable
types of MALT include gut-associated lymphoid tissue (GALT) and bronchus-associated
lymphoid tissue (BALT), both of which are well-characterized.\

MALT
Image source: Kuby Immunology 7th Edition
3.1.1. Gut-Associated Lymphoid Tissue (GALT):
GALT, found in the gastrointestinal tract, is one of the best-characterized types of
MALT. It comprises Peyer's patches and isolated follicles in the tissue beneath the colon's
mucosa. Peyer's patches are lymphocyte aggregates forming primary lymphoid follicles (30-40
follicles with centrally located B cells surrounded by T cells and macrophages) that can develop
into secondary follicles with germinal centers upon antigen activation. These patches have
efferent lymphatics draining lymph into mesenteric lymph nodes but lack afferent lymphatics.
Peyer's patches are covered by specialized lympho-epithelium made up of microfold (M)
cells in small regions called inductive sites. M cells are flattened epithelial cells without the
microvilli typical of mucous epithelium, and they have deep pockets filled with clusters of B
cells, T cells, and macrophages. The epithelial cells of mucous membranes are crucial in
promoting the immune response by encountering antigens in the gut. M cells transport antigens
across the mucous membrane, activating B cells within lymphoid follicles. These activated B
cells then differentiate into IgA-secreting plasma cells. The antibodies are transported across
epithelial cells and released as secretory IgA into the lumen, where they interact with antigens
present. This is the site where secretory IgA is concentrated.

a) M Cell b) Transport of IgA

Image source: Kuby Immunology 7th edition
3.1.2. Bronchus-Associated Lymphoid Tissue (BALT):
Another well-characterized type of MALT is BALT, which is structurally similar to
GALT. BALT consists of large collections of lymphocytes, mainly B cells, organized into
aggregates and follicles with a few germinal centers. These are located primarily along the main
bronchi in the lungs. The epithelium covering BALT follicles lacks goblet cells and cilia. The M
cells of BALT follicles are structurally similar to the intestinal M cells of GALT. BALT contains
a complex network of capillaries, arterioles, venules, and efferent lymphatics, suggesting that
BALT may sample antigens not only from the lungs but also from the systemic circulation.
3.2. Cutaneous-Associated Lymphoid Tissue (CALT):
The skin serves as a crucial barrier against external environments due to its large surface
area, playing an essential role in the body's nonspecific (innate) defenses. The outer layer of the
skin, the epidermis, is primarily composed of specialized epithelial cells called keratinocytes.
These cells secrete cytokines that may induce local inflammatory reactions and can be induced to
express class II MHC molecules, functioning as antigen-presenting cells (APCs).
Within the epidermis, a type of dendritic cell known as Langerhans cells is scattered
among the epithelial-cell matrix. Langerhans cells capture antigens in the epidermis through
phagocytosis or endocytosis. They then migrate to regional lymph nodes and differentiate into
interdigitating dendritic cells. These cells express high levels of class II MHC molecules and act
as potent activators of naive T helper (TH) cells by presenting antigens to them.

CALT
Image source: Kuby Immunology 7th edition
3.3. Intraepithelial Lymphocytes (IELs):
The outer mucosal epithelial layer contains intraepithelial lymphocytes (IELs), many of
which are T cells with unique receptors (γδT-cell receptors or γδTCRs) that have limited antigen
diversity and are well-positioned to encounter antigens entering through the intestinal mucous
epithelium. A large number of lymphocytes are associated with the epithelial surfaces of the
body, especially in the reproductive tract, lungs, and skin. The dermal layer of the skin contains
CD4+ and CD8+ T cells, as well as macrophages. Most of these dermal T cells are either
previously activated cells or memory cells. These collections of lymphoid cells play a crucial
role in developing mucosal immunity, contributing to both local and systemic immune responses
to antigens at the body surface.
 Innate and adaptive immunity
The immune system is a complex network of immune cells and proteins like cytokines
working together to fight off pathogens such as bacteria, viruses, fungi, parasitic worms, and
abnormal cells. These cells have a remarkable ability to distinguish between self and non-self
molecules, although this ability can be compromised in conditions such as autoimmune diseases.
The differentiation between self and non-self is based on the Major Histocompatibility Complex
(MHC) proteins present on the surface of all body cells, except for identical twins.
There are two types of MHC proteins: MHC I and MHC II. MHC I is responsible for
differentiating the body’s own cells from foreign cells or pathogens and displaying antigens on the
cell surface. MHC II, found in antigen-presenting cells (APCs) along with MHC I, displays
phagocytosed microbes on the cell surface. MHC proteins are absent in red blood cells.
The body has two types of responses to invaders or antigens: Innate (Natural/Non-specific) and
Acquired (Adaptive/Specific).
1. Innate Responses:
 These occur to the same extent regardless of how many times the pathogen is
exposed.
 They employ phagocytic cells such as neutrophils, monocytes, macrophages, and
Natural Killer (NK) cells.
 They respond immediately to foreign attacks.
 They fight against all invaders, hence termed non-specific.
 They comprise the first and second lines of defense.
2. Acquired Responses:
 These responses improve with repeated subsequent exposures to foreign particles.
 They involve antigen-specific B and T cells or antigen-presenting cells (APCs).
 They take a longer time to react compared to innate responses.
 They fight against specific types of invaders.
 They include the third line of defense.
 General principles and mechanism of innate immunity
Image
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169770300515/General-principles-of-the-innate-immune-response-The-innate-immune-system-is-
a.png

Basically, there are three types of lines of defense:

1. First line of defense
2. Second line of defense
3. Third line of defense
 Image source: https://msgallagherlhs.weebly.com/uploads/3/8/6/9/38694679/1344533_orig.png

First Line of Defense

The first line of defense, also known as the outside defense system, comprises physical,
chemical, and biological defenses that protect against the invasion of pathogens. These defenses do
not function independently and often overlap in their roles.
Physical Defenses
Physical defenses include barriers and mechanical defenses that block the entry points of pathogens,
such as intact skin and mucus.
1. Skin:
 The skin has three layers: the epidermis, the dermis, and the hypodermis.
 The topmost layer, the epidermis, is packed with keratin and dead skin cells, which
are frequently shed and replaced.
 Keratin is highly water-resistant and mechanically tough, resisting microbial growth.
2. Nasal Hairs and Cilia:
 Nasal hairs filter air contaminated with microbes, dust, and dirt.
 Microscopic cilia lining the respiratory tract sweep mucus and trapped particles
toward body openings for removal.
3. Mucous Membranes:
 Mucous membranes lining the respiratory, urinary, and reproductive tracts produce
mucus, a slimy substance that traps foreign particles.
  These particles are directed out of the body through mechanical actions such as
shedding, coughing, peristalsis, and the flushing of bodily fluids (e.g., urination,
tears).
These coordinated defenses form a robust first line against pathogenic invaders, keeping the
body safeguarded from external threats.

Chemical Defenses
Chemical defenses in the body include chemicals and enzymes in body fluids, plasma
protein mediators, cytokines, antimicrobial peptides, and inflammation-eliciting mediators. These
elements destroy pathogens on the outer body surface, at body openings, and along inner body
linings.

1. Body Fluids:
 Sweat, Tears, Mucus, and Saliva: Contain enzymes like lysozyme that break down
bacterial cell walls, killing the bacteria. Secretory IgA similarly attacks bacterial cell
walls.
2. Antimicrobial Peptides (AMPs):
 Include dermcidin, cathelicidin, defensins, histatins, and bacteriocins. These AMPs
are produced in response to pathogens on the skin and help destroy them.
3. Cerumen (Ear Wax):
 Contains fatty acids that lower the pH to between 3 and 5, protecting the auditory
canal from foreign particles like microbes.
4. Gastric Juice:
 With a highly acidic pH of 2-3, it destroys pathogens that enter the stomach through
the oral cavity or nasal tract.
5. Urine Flow:
 Acidic in pH, it helps kill microbes and flushes them out of the urethra.
6. Serum:
 Contains unsaturated fatty acids that reduce water loss and inhibit microbial growth.
However, it also contains certain compounds that provide nutrition for some
microbes.
These chemical defenses work together to protect the body from a wide range of pathogens,
ensuring a robust defense against infections.
 Chemical defenses of innate immune system
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Biological Defenses
Biological defenses are provided by living microorganisms that are friendly and beneficial.
These are known as resident natural flora and are found on our skin, in our bowel, and in other
places such as the mouth, gut, and reproductive parts. These microorganisms play a crucial role in
protecting our bodies from pathogenic invaders.

How Resident Natural Flora Protect Us?

1. Creating an Acidic Environment:
 Through the fermentation of sugars to acids, these microorganisms create an acidic
environment that is hostile to many pathogens.
2. Occupying Binding Sites:
 By colonizing available cellular binding sites, they prevent pathogens from adhering
to and colonizing our bodies.
3. Competing for Nutrients:
 They compete with pathogens for available nutrients, making it difficult for harmful
microorganisms to thrive.
Additionally, the resident normal microbiota contributes to our chemical defenses by
producing bacteriocins. Bacteriocins are antibacterial proteins that inhibit the growth of or kill other
bacteria. This helps maintain the balance of good bacteria and prevents the overgrowth of harmful
bacteria.These multifaceted biological defenses form an essential part of our immune system,
working synergistically with physical and chemical defenses to keep us healthy.

Second Line of Defense

When the first line of defense is broken, the second line of defense within our body ‘kicks’
in and gets activated i.e. when the pathogens successfully win the battle against barriers from the
first line of defense, they are next encountered by the second line. This mechanism involves the
participation of mainly immune cells mainly white blood cells (leucocytes) such as phagocytes, NK
cells, dendritic cells, mast cells, and complement proteins to recognize and eliminate any non-
specific pathogen entered into the body. There is no immunological memory.This internal defense
mechanism is crucial in recognizing and eliminating non-specific pathogens that have successfully
bypassed the initial barriers.

Key Components and Functions:

1. Immune Cells (Leucocytes):
a) Phagocytes: These include neutrophils, monocytes, and macrophages. They engulf
and digest pathogens and debris.Macrophages are long-lived phagocytic cells that
reside in tissues and engulf pathogens, debris, and apoptotic cells. Secrete cytokines
and present antigens to T cells, linking innate and adaptive immunity.
b) Natural Killer (NK) Cells: These cells target and destroy infected or abnormal cells
by inducing apoptosis (programmed cell death).Natural killer (NK) cells are
cytotoxic lymphocytes that recognize and kill virus-infected and tumor cells. Secrete
cytokines (interferon-gamma) that activate macrophages and enhance antiviral
responses.
c) Dendritic Cells: They capture and present antigens to T cells, initiating an immune
response.Mature upon encountering pathogens and migrate to lymph nodes to initiate
adaptive immune responses
d) Mast Cells: Mast cells are tissue-resident cells that release histamine and other
mediators in allergic responses. Involved in allergic reactions and also play a role in
wound healing and defense against pathogens.
 Cells of the Innate Immune System
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-of-innate-and-adaptive-immune-system-cells-Innate-immune-responses-are.jpg

2. Complement Proteins:
a) These proteins circulate in the blood and, upon activation, enhance the ability of
antibodies and phagocytic cells to clear microbes and damaged cells from the
organism.
b) They promote inflammation and attack the pathogen's cell membrane, leading to
its destruction.

Basic Mechanism of the Second Line of Defense

When a pathogen invades at the entry site, neutrophils are the first to respond by
engulfing and destroying the invaders. If the pathogens evade the neutrophils, macrophages and
dendritic cells step in to phagocytose the invaders and present antigens to T cells. The second line of
defense includes various internal mechanisms like phagocytosis, NK cells, inflammatory response,
fever, and the complement system.
 1. Recognition:
a. Immune cells recognize pathogens through pattern recognition receptors (PRRs) that
identify pathogen-associated molecular patterns (PAMPs) on the surface of microbes.
2. Response:
a. Once a pathogen is recognized, immune cells and proteins work together to eliminate
the invader through various mechanisms such as phagocytosis, cytotoxicity, and the
release of antimicrobial substances.

Mechanism of Innate Immunity

Image source: https://microbenotes.com/wp-content/uploads/2022/05/Innate-Immunity.jpeg

Key Mechanisms:
1. Phagocytosis:
 Neutrophils, macrophages, and dendritic cells engulf and digest pathogens.
2. Natural Killer (NK) Cells:
 These cells detect and induce apoptosis in virally infected or cancerous cells.
3. Inflammatory Response:
 Immune and infected cells release cytokines that amplify cytokine secretion,
triggering an inflammatory response.
 This response includes capillary dilation and increased permeability of the capillary
wall.
 Macrophages help resolve the inflamed site by clearing up cellular debris.
 4. Fever:
 Cytokines increase core body temperature, resulting in fever.
 This elevated temperature inhibits microbial growth and accelerates recovery and
repair processes.
5. Complement System:
 Complement proteins in the blood serum are attracted to pathogens tagged by the
adaptive immune system.
 A cascade of complement protein binding coats the pathogens, marking them for
detection by phagocytes and facilitating their digestion through phagocytosis.
These coordinated responses ensure that pathogens are effectively recognized and eliminated,
maintaining the body's defense against infections.
Lack of Immunological Memory:
Unlike the adaptive immune response, the second line of defense does not generate
immunological memory. This means it does not "remember" previous encounters with specific
pathogens, and the response is the same each time a pathogen is encountered.This second line of
defense is a critical part of our body's innate immune response, providing a rapid, non-specific
reaction to infections and helping to contain and eliminate pathogens before they can cause
significant harm.

Recognition of Pathogens by Innate Immune System

1) Innate immune cells express pattern recognition receptors (PRRs) that detect pathogen-
associated molecular patterns (PAMPs)
2) PAMPs are conserved molecular structures essential for pathogen survival and absent in host
cells (lipopolysaccharide, peptidoglycan, double-stranded RNA)
3) Toll-like receptors (TLRs) are a major class of PRRs expressed on cell surface or in
endosomes
a. Different TLRs recognize specific PAMPs: TLR4 (lipopolysaccharide), TLR3
(double-stranded RNA), TLR9 (CpG DNA)
4) Other PRRs include C-type lectin receptors (CLRs), NOD-like receptors (NLRs), and RIG-
I-like receptors (RLRs)
5) PRR engagement activates signaling cascades that lead to production of cytokines,
chemokines, and antimicrobial molecules
6) Innate immune cells also express receptors for opsonins (complement, antibodies) that
enhance phagocytosis
 7) Natural killer (NK) cells express activating and inhibitory receptors that regulate their
cytotoxic activity
a. Activating receptors (NKG2D) recognize stress-induced ligands on infected or tumor
cells
b. Inhibitory receptors (KIRs) recognize self MHC class I molecules and prevent NK
cell activation

Recognition of Pathogens by Innate Immune System

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Innate Immune Responses of different types of immune cells:

1) Phagocytosis is the process by which phagocytic cells (neutrophils, macrophages) engulf
and destroy pathogens
a. Involves recognition of PAMPs or opsonized pathogens, actin polymerization, and
formation of a phagosome
b. Phagosome fuses with lysosomes containing antimicrobial enzymes and reactive
oxygen species
2) Neutrophils undergo oxidative burst, generating reactive oxygen species (superoxide,
hydrogen peroxide) that kill pathogens
3) Neutrophils and other granulocytes release granules containing antimicrobial enzymes
(myeloperoxidase, defensins) and proteases
4) Natural killer (NK) cells release cytotoxic granules containing perforin and granzymes that
induce apoptosis in target cells
5) Innate immune cells secrete cytokines and chemokines that regulate immune responses and
recruit additional immune cells
a. Macrophages and dendritic cells produce IL-1, IL-6, IL-12, and TNF-alpha
b. Neutrophils and epithelial cells produce IL-8, a potent neutrophil chemoattractant
6) Complement system enhances phagocytosis (opsonization), recruits inflammatory cells
(anaphylatoxins), and directly lyses pathogens (membrane attack complex).
7) Interferons (type I and II) are cytokines that induce an antiviral state in infected and
neighboring cells
a. Type I interferons (IFN-alpha, IFN-beta) are produced by most cells in response to
viral infection
b. Type II interferon (IFN-gamma) is produced by NK cells and T cells and activates
macrophages
 Basic Properties and types of Antigens

Antigens are molecules or molecular structures that are foreign to the body and generally
induce an immune reaction by triggering the production of antibodies against them. As the
Antigens are substances that do not belong to the body, they are perceived as foreign invaders.
Antigens are typically defined by their ability to induce an immune response, but not all antigens
have the capacity to trigger such a response. The antigens that do induce a response are known as
immunogens.Antigens, indicated by the term 'Ag', can come in various forms including pollen,
viruses, chemicals, and bacteria.

The ability of antigens to elicit an immune response depends on specific regions called
antigenic determinants or epitopes. These determinants bind to receptor molecules with
complementary structures on immune cells to trigger a response.The concept of an antigen is
rooted in the body's ability to distinguish between its own components and foreign particles. This
distinction is crucial for an effective immune response.In response to antigens, the body produces
antibodies that specifically target and act against these foreign substances.Most antigens in
humans are proteins, peptides, or polysaccharides. However, lipids and nucleic acids can also
function as antigens when combined with proteins or polysaccharides.

Antigens might also be intentionally introduced into the body in the form of vaccines to
stimulate the adaptive immune system. This controlled exposure helps the body develop
immunity against specific pathogens.The presence of antigenic determinants on antigens allows
them to bind to receptor molecules on immune cells, leading to the activation of the immune
response. This process ensures that the body can identify and combat a wide array of pathogens
effectively.These insights highlight the crucial role of antigens in the immune system,
underpinning the body's defense mechanisms against a variety of foreign invaders.

Structure of Antigen
The structure of an antigen is characterized by its ability to bind to the antigen-binding site
of an antibody. Antibodies can differentiate between different antigens based on the unique
molecular structures present on the surface of the antigen.The molecular structure of an antigen
is characterized by its ability to bind to the antigen-binding site of an antibody.Most antigens are
proteins or polysaccharides. These can include coats, capsules, flagella, toxins, and fimbriae of
bacteria, viruses, or other microorganisms. Besides, secretions and other chemicals of the same
nature can also act as antigens.Lipids and nucleic acids of these microorganisms are only
antigenic when these are combined with proteins or polysaccharides.
Ideal immunogens are neither too large nor too small, with a molecular weight range of 8-
10K Da. Very small molecules can be made immunogenic by attaching them to larger molecules.
Antigens can be proteins, polysaccharides, lipids, or nucleic acids. The presence of aromatic
radicals is thought to enhance a molecule's antigenicity. Peptide antigens can generate a strong
immune response if they contain a high number of hydrophilic and charged amino acids such as
lysine, glutamine, glutamic acid, asparagine, and aspartic acid. These antigens should be
structurally different from the host's biomolecules and must interact easily with immune cells to
trigger an immune response.
The structure of antigens might be different depending on the nature of the antigen, their
size, and immunogenicity.All immunogenic antigens have a specific structural component called
epitope or antigenic determinant.The number of epitopes differs in different antigens and
determines the number of antibodies a single antigen can bind to.The region on antibodies that
interacts with antigens is called a paratope. It has been established that the structure of epitope
and paratope can be defined with a lock and key metaphor as the structures are specific and fit
with one another.
Apart from the chemical form, the physical form of a molecule can also influence its
immunogenicity. Particulate matter has been found to provoke a stronger immune response
compared to its soluble form. Interestingly, the denatured form of a biomolecule can elicit a
greater immune response than its native structure. Inert substances, such as dust, which cannot be
degraded by immune cells, tend to cause allergic reactions rather than antibody production.
Some of the structural Components of Antigens are:
1. Proteins and Polysaccharides:
 Most antigens are proteins or polysaccharides. These can include components
such as coats, capsules, flagella, toxins, and fimbriae of bacteria, viruses, or other
microorganisms.
 Secretions and other chemicals of a similar nature can also act as antigens.
2. Lipids and Nucleic Acids:
 While lipids and nucleic acids are not typically antigenic on their own, they can
become antigenic when combined with proteins or polysaccharides.
 3. Epitopes (Antigenic Determinants):
 All immunogenic antigens possess specific structural components known as
epitopes or antigenic determinants.
 The number of epitopes on an antigen varies, and this determines the number of
antibodies that can bind to a single antigen.
 Epitopes bind to receptor molecules with complementary structures on immune
cells to elicit an immune response.
4. Paratopes:
 The region on antibodies that interacts with antigens is called a paratope.
 The relationship between an epitope and a paratope can be likened to a lock and
key metaphor, where the structures are highly specific and fit together precisely.

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Properties of Antigens:
Antigens possess various properties that determine their immunogenicity, making them
crucial for understanding the immune response against them. Since these properties influence
immunogenicity, they are essential for identifying an effective antigen. Some of the key
properties of antigens include:
1. Foreign Nature
 Antigens that trigger an immune response in the host are foreign to the recipient's
body.
 The host body identifies the antigen as different from its normal components.
 The immunogenicity of the antigen increases with the degree of foreignness. For
biological antigens, foreignness increases with the phylogenetic gap between
species.
 However, exceptions exist, such as certain proteins within the host that may
induce an immune response, known as autoantigens.
 Additionally, proteins and molecules from other species might not trigger an
immune response if they lack specific antigenic determinants or epitopes.
2. Chemical nature of antigens:

 The most effective and commonly found antigens are proteins, followed by
polysaccharides.
 Other molecules, such as lipids and nucleic acids, can also serve as antigens when
they form complexes with proteins and polysaccharides.
 For proteins to act as antigens, they should have immunogenic regions containing at
least 30% of amino acids like lysine, glutamine, arginine, glutamic acid, asparagine,
and aspartic acid, along with a high number of hydrophilic or charged groups.
 The level of immunogenicity increases with the heterogeneity of the molecules.
Homopolymers are generally less immunogenic than heteropolymers.

3. Molecular Size:
 The molecular size of the antigens is also crucial in the immunogenicity of the
molecules.
 It has been established that antigens should have a minimum size of greater than 5000
Da before they can be considered immunogenic.
 However, low molecular weight substances can demonstrate immunogenicity when
coupled with large-sized carriers.
 The low molecular weight substances are termed haptens that are considered ‘partial
antigens’ with at least one antigenic determinant.
4. Molecular Rigidity and Complexity:
 The rigidity and complexity of molecules are essential factors that determine
immunogenicity.
 In general, rigid molecules are good antigens as they can raise antibodies to certain
structures when compared to the less rigid ones.
 The complexity of the structure is also an essential factor as a peptide antigen with a
repeating unit of a single amino acid is less immunogenic than a molecule with two or
more repeating amino acids units.

5. Antigenic Determinants and Cross-reactivity:

 Antigenic determinants, also known as epitopes, are specific regions on an antigen

molecule that interact with antibodies.
 Typically, antigens that have two or more antigenic determinants are capable of
inducing antibody production.
 In contrast, smaller antigens usually cannot trigger antibody production, as they may
lack multiple antigenic determinants.
 Cross-reactivity is another important aspect where antibodies produced in response to
one antigen can also recognize and react with a different antigen.

6. Genetic makeup of the host:

 can lead to different responses to the same antigen in two strains of the same animal
species.
 This variation arises from differences in the genes that govern the immune response,
such as the diversity in MHC alleles.

7. Dosage, route, and timing of antigen administration:

 The magnitude of the immune response is significantly influenced by the amount of

antigen administered.
 Consequently, optimizing the immune response involves adjusting the dosage, route,
and timing of antigen administration.
  Antigens can be administered through various routes, including intravenous,
subcutaneous, and intradermal methods, to provoke an immune response.
 Subcutaneous and intradermal routes elicit a stronger response compared to the
intravenous route
 . When administered intravenously, the antigen first travels to the spleen, whereas
antigens introduced subcutaneously first migrate to the local lymph nodes.
 Certain molecules that are not inherently immunogenic can enhance the immune
response when combined with other substances, known as adjuvants. An example of an
adjuvant is aluminum hydroxide.
 Adjuvants are administered alongwith an antigen to assist the immune system in
producing antibodies that target the antigen.
 Combining the antigen with an adjuvant an agent that enhances immunogenicity by
aiding the delivery of antigens to Antigen-Presenting Cells (APCs) can further boost the
immune response.

8. Degradability of antigens:

 Antigens that are easily engulfed by phagocytes typically trigger a stronger immune
response.
 Molecules with higher biodegradability are more effective as antigens. This is because
most antigens must be phagocytosed, processed, and presented to helper T cells by
antigen-presenting cells (APCs) to initiate an immune response.
 Conversely, non-biodegradable substances like dust particles tend to cause allergic
reactions rather than an immunogenic response.

Types of Antigens:
Antigens can be categorized in various ways, with common classifications based on their origin
and immunogenicity.
1. Types of Antigens Based on Their Origin
Antigens can be classified into the following groups based on their origin:
a. Exogenous Antigens
 Exogenous antigens are foreign substances that enter an organism's body from the
external environment. They can penetrate the body through ingestion, inhalation, or injection,
and are considered non-self-antigens. These antigens are found in body fluids and extracellular
spaces and are associated with MHC Class II molecules. Examples of exogenous antigens
include bacteria, viruses, allergens like pollen, and toxic foods.The uptake of exogenous antigens
is primarily mediated by phagocytosis via Antigen Processing Cells (APCs) like macrophages,
dendritic cells, etc. Some antigens initially enter as exogenous antigens but can become
endogenous, such as intracellular viruses.

Exogenous antigens
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b. Endogenous Antigens
Endogenous antigens are those that are generated within the cells of the host. These
antigens can be produced due to normal cell metabolism or as a result of infections by
intracellular pathogens such as viruses, bacteria, and parasites. Endogenous antigens may also be
the cells of the body or fragments, compounds, or antigenic products of metabolism.Endogenous
antigens might result in autoimmune diseases as the host immune system detects its own cells
and particles as immunogenic.
 Endogenous antigens are processed by the macrophages and presented on the cell surface
by Major Histocompatibility Complex (MHC) class I molecules, which are then recognized by
cytotoxic T cells (CD8+ T cells).

Endogenous antigens
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419679259024475638/

c. Autoantigens
Autoantigens are proteins or protein complexes from the host that are mistakenly targeted
by the host's own immune system, leading to autoimmune diseases. This misdirected immune
response can be harmful, as the immune system attacks the body's own cells. The loss of
immunological tolerance to these antigens is influenced by genetic and environmental factors.
d. Tumor Antigens (Neoantigens)
Tumor antigens, or neoantigens, are displayed by Major Histocompatibility Complex
(MHC) I and II on the surfaces of tumor cells. These antigens arise from tumor-specific
mutations during the malignant transformation of normal cells. Typically, tumor cells develop
mechanisms to avoid antigen presentation and immune detection, preventing a robust immune
response.
e. Native Antigens
Native antigens are those that have not been processed by antigen-presenting cells
(APCs), which means that immune cells like T-cells cannot recognize them. However, B-cells
can be activated by these unprocessed antigens independently.
2. Types of Antigens Based on Immune Response:
Antigens can be categorized into two main groups based on their ability to elicit an immune
response:
a. Complete Antigens/Immunogens
Complete antigens, or immunogens, are capable of provoking a specific immune
response on their own. These antigens are typically proteins, peptides, or polysaccharides with a
high molecular weight, generally exceeding 10,000 Da.Some features of the complete antigens
are characteristics of complete antigens:
 Self-Sufficient: These antigens can stimulate an immune response without the need for

any carrier particles.

 Molecular Composition: They are usually composed of proteins, peptides, or

polysaccharides.
 Antigenic Determinants: Complete antigens have multiple antigenic determinant sites,

also known as epitopes, which are recognized by antibodies or T-cell receptors.

 Complete antigens are crucial in triggering the immune system to produce antibodies or

activate T cells, thereby playing a significant role in the body's defense mechanisms.
b. Incomplete Antigens/Haptens
Incomplete antigens, or haptens, are unable to generate an immune response by
themselves. They are usually non-protein substances that require a carrier molecule to form a
complete antigen. Haptens have a low molecular weight, generally less than 10,000 Da, and
possess fewer antigenic determinant sites. The carrier molecule attached to the hapten, usually a
protein or polysaccharide, is considered non-antigenic.
 They are often non-protein substances, such as drugs, chemicals, or small organic
compounds.
 Haptens possess fewer antigenic determinant sites compared to complete antigens.
 The immune system recognizes the conjugate formed by the hapten and the carrier
molecule.
  Once bound to a carrier, haptens can induce the production of specific antibodies
against the hapten.
 Haptens are often used in research and vaccine development to create artificial
antigens.
 Some drug allergies are caused by the immune system reacting to drug molecules
acting as haptens.
 Haptens can form epitopes, the specific parts of an antigen recognized by antibodies
or T-cell receptors, when attached to a carrier.
 The immune response is directed against both the hapten and the carrier molecule.
 Conjugation to a carrier enhances the immunogenicity of haptens.
 Common examples of haptens include penicillin, urushiol (the compound in poison
ivy), and certain cosmetic and food additives.
 Haptens are valuable tools in immunological research for studying immune
responses.
 Haptens are used in diagnostic tests to detect specific antibodies in the blood, such as
in allergy testing.

Mechanism and Role of Antigen and Antibody Interaction in Immunology:

1. Antigen Presentation and Recognition
 Antigen Presentation: Dendritic cells, macrophages, and B cells capture antigens and

present them on their surface using molecules called MHC (Major Histocompatibility
Complex). T cells recognize these antigen-MHC complexes.
 B Cell Receptors: B cells have membrane-bound antibodies (B cell receptors or BCRs)

that specifically recognize and bind to antigens. Each B cell has a unique BCR, tailored
to recognize a specific antigen.
2. Clonal Selection and Expansion
 Activation: When a B cell's receptor binds to an antigen, the B cell is activated. Helper T

cells (specifically T_H cells) often assist in this activation by releasing cytokines.
 Clonal Expansion: The activated B cell proliferates (clonal expansion) and differentiates

into plasma cells and memory B cells. Plasma cells produce large quantities of antibodies
specific to the antigen.
3. Antibody-Antigen Binding
  Variable Region: The variable regions of the antibody (comprising the tips of the "Y"

shape) are highly specific to the antigen. They form a unique binding site that fits the
antigen's epitope (the specific part of the antigen recognized by the antibody).
 Formation of Immune Complex: The binding of an antigen to an antibody forms an

antigen-antibody complex. This binding is highly specific, similar to a lock and key
mechanism.
4. Effector Functions of Antibodies
 Neutralization: Antibodies can neutralize pathogens by binding to them and blocking

their ability to infect cells. For example, antibodies can prevent viruses from attaching to
host cells.
 Opsonization: Antibodies coat the surface of pathogens, enhancing their recognition and

ingestion by phagocytes like macrophages and neutrophils.

 Agglutination and Precipitation: Antibodies can cause antigens to clump together

(agglutination) or to precipitate out of solution, making it easier for phagocytes to engulf

them.
 Complement Activation: The Fc region of bound antibodies can activate the

complement system. This leads to the formation of the membrane attack complex (MAC)
which can lyse (burst) the pathogen.
 Antibody-Dependent Cellular Cytotoxicity (ADCC): Natural killer (NK) cells

recognize and bind to the Fc region of antibodies attached to infected cells or pathogens.
The NK cells then release cytotoxic molecules to kill the target.
5. Memory Formation
 Memory B Cells: After the initial immune response, some B cells differentiate into

memory B cells. These cells persist in the body and can quickly result in a strong
response if the same antigen is encountered again in the future.

Understanding the complex structure of antigens and their interactions with antibodies is
essential for understanding how the immune system identifies and combats foreign invaders,
ensuring effective protection and immune surveillance.
 Antigen – Antibody Interaction
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 B and T Cell Epitopes, Paratopes
B Cell Epitopes: Immunity Defenders
B cell epitopes, or antigenic determinants, are pivotal elements of the immune system.
They are specific regions of an antigen that are recognized by B cells, leading to an immune
response. The identification and characterization of B cell epitopes are critical for vaccine
development, diagnostics, and therapeutic interventions. B cell epitopes are part of the antigen
that binds to the B cell receptor (BCR) on the surface of B cells. This binding triggers a
cascade of events, leading to the activation, proliferation, and differentiation of B cells into
plasma cells that secrete antibodies. These antibodies specifically target and neutralize the
pathogen. The identification and characterization of B cell epitopes are critical for vaccine
development, diagnostics, and therapeutic interventions.

Epitopes:

Epitopes, also known as antigenic determinants, are specific parts of an antigen that
are recognized and bound by immune system components such as antibodies, B cell receptors
(BCRs), or T cell receptors (TCRs). They are critical for the immune system's ability to
identify and respond to pathogens.

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Detailed Structure of B Cell Epitopes:

The structure of B cell epitopes is fundamental to their function in the immune

response. They are categorized into two main types: linear epitopes and conformational
epitopes. Each type has unique structural characteristics that influence how they are
recognized by antibodies.

1. Linear Epitopes

⬥ Primary Structure: Linear epitopes are composed of a continuous sequence of amino

acids within the protein's primary structure. This sequence forms a specific linear
segment that antibodies recognize.
⬥ Recognition: These epitopes are recognized by antibodies based on their linear amino

acid sequence. The antibody's binding site, or paratope, interacts with this specific
sequence.
⬥ Stability: Linear epitopes remain recognizable to antibodies even when the protein is

denatured, as their sequence is unaltered. This property is advantageous for diagnostic

assays like ELISAs, which often use denatured proteins.
⬥ Example: In the case of a viral protein, a linear epitope might be a segment like -A-G-

L-P-Q-T-R-, where each letter represents an amino acid.

2. Conformational (Discontinuous) Epitopes

⬥ Tertiary Structure: Conformational epitopes are composed of amino acids that are

not sequentially adjacent but come together in the protein's three-dimensional folded
structure. This spatial arrangement forms a unique shape recognized by antibodies.
⬥ Recognition: These epitopes are recognized by antibodies based on the protein's three-

dimensional structure. The antibody's paratope fits the specific shape of the
conformational epitope.
⬥ Stability: Conformational epitopes are dependent on the protein's native structure.

Denaturation or alteration of the protein's folding can disrupt the epitope, making it
unrecognizable to antibodies.
 ⬥ Example: In a folded protein, a conformational epitope might involve amino acids

from different parts of the primary sequence coming together in a specific spatial
arrangement

Conformational epitope: Amino acid residues located at different parts in the

polypeptide chain. Linear epitope: Amino acid residues are adjacent in the polypeptide
chain
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Importance of B Cell Epitopes in the Immune Response:

B cell epitopes, also known as antigenic determinants, play a crucial role in the
immune response. They are specific regions on antigens that are recognized by B cells,
triggering a series of immune reactions. Understanding the importance of B cell epitopes is
vital for vaccine development, disease diagnostics, and therapeutic interventions.

1. Recognition and Binding

The primary function of B cell epitopes is to be recognized and bound by B cell

receptors (BCRs) on the surface of B cells. This recognition is highly specific, allowing B
cells to distinguish between different antigens. The binding of an epitope to a BCR initiates
the activation of B cells, leading to their proliferation and differentiation into plasma cells.

2. Antibody Production

Once activated, B cells differentiate into plasma cells that produce antibodies specific
to the recognized epitope. These antibodies circulate in the bloodstream and lymphatic
system, seeking out and neutralizing pathogens. The ability of B cells to produce antibodies
against specific epitopes is essential for the body to mount an effective immune response
against infections.

3. Immune Memory

B cell epitopes are also critical for the formation of immune memory. Memory B cells
are generated during the primary immune response and persist long-term in the body. Upon
re-exposure to the same antigen, these memory B cells rapidly respond by producing large
quantities of antibodies. This mechanism provides long-lasting protection against recurrent
infections and is the basis for the effectiveness of vaccines.

4. Vaccine Development

The identification and characterization of B cell epitopes are fundamental in vaccine

development. By targeting specific epitopes on pathogens, vaccines can elicit strong and
protective immune responses. Epitope-based vaccines focus on the most immunogenic regions
of an antigen, improving the efficiency and safety of vaccines. Techniques such as epitope
mapping and computational prediction are used to identify these critical regions.

5. Diagnostic Applications

In diagnostics, B cell epitopes are utilized to detect the presence of specific antibodies
in a patient's blood. This can indicate exposure to a pathogen or the effectiveness of a
vaccination. Diagnostic assays, such as enzyme-linked immunosorbent assays (ELISAs),
often employ synthetic peptides representing B cell epitopes to capture and measure
antibodies.

6. Therapeutic Interventions

Monoclonal antibodies targeting specific B cell epitopes have been developed for
various therapeutic applications. These monoclonal antibodies can neutralize pathogens,
inhibit disease progression, and modulate immune responses. They are used in the treatment
of diseases such as cancer, autoimmune disorders, and infectious diseases.
Functions of B Cell Epitopes:

1. Recognition by B Cells

B cell epitopes are specific regions on antigens that are recognized and bound by B
cell receptors (BCRs) on the surface of B cells. This recognition is incredibly precise, as
BCRs have unique binding sites that match the epitopes. This specificity ensures that B cells
can accurately identify and bind to foreign pathogens.

2. Initiation of Immune Response

The binding of an epitope to a BCR triggers a signaling cascade within the B cell,
leading to its activation. This process involves several steps:

 Activation: The B cell receptor, upon binding to the epitope, sends signals to the B

cell's interior, activating it.

 Proliferation: The activated B cell undergoes rapid division, producing a large

number of identical B cells, all capable of recognizing the same epitope.

 Differentiation: These B cells then differentiate into two types of cells: plasma cells

and memory B cells.

3. Antibody Production

Plasma cells are the differentiated B cells that produce and secrete antibodies specific
to the epitope. These antibodies circulate throughout the body, binding to the same epitopes
on pathogens. This binding marks the pathogens for destruction by various immune
mechanisms. The specificity of antibodies ensures that they target only the harmful invaders
without affecting the body's own cells.
 Role of b cell epitope in antibody production
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4. Formation of Immune Memory

During the immune response, some of the activated B cells differentiate into memory
B cells. These cells persist in the body long after the initial infection has been cleared. Upon
re-exposure to the same antigen, memory B cells quickly recognize and respond to the
specific epitopes, producing antibodies rapidly and in large quantities. This mechanism
provides long-term immunity and is the basis for the effectiveness of vaccines.

5. Enhancement of Phagocytosis

Antibodies bound to epitopes on pathogens can enhance the process of phagocytosis,

in which phagocytic cells like macrophages and neutrophils engulf and digest the pathogens.
This enhancement occurs through a process called opsonization, where antibodies coat the
surface of the pathogen, making it easier for phagocytic cells to recognize and ingest them.
6. Complement Activation

Antibody-bound epitopes can activate the complement system, a group of proteins that
work together to destroy pathogens. This activation triggers a series of reactions that lead to
the formation of the membrane attack complex (MAC). The MAC forms pores in the
membrane of the pathogen, leading to its lysis and destruction.

Role of B cell epitopes in Immune response mechanism

T Cell Epitopes
T cell epitopes, also known as T cell antigenic determinants, are specific regions of
an antigen that are recognized by T cell receptors (TCRs) on the surface of T cells. These
epitopes play a critical role in the immune response, particularly in the activation and function
of T cells.
Structure of T Cell Epitopes:
T cell epitopes are short peptides presented by major histocompatibility complex
(MHC) molecules on the surface of cells. Their structure and presentation are crucial for the
activation of T cells.
1. Composition of T Cell Epitopes: T cell epitopes are typically composed of short peptide
sequences derived from proteins. These peptides are usually 8-25 amino acids in length. Their
structure is essential for binding to MHC molecules and recognition by T cell receptors
(TCRs).
2. MHC Class I and Class II Presentation MHC Class I Molecules: These molecules present
peptides to CD8+ cytotoxic T cells. The peptides are typically 8-10 amino acids long and
derived from intracellular proteins.
The MHC class I molecule has a binding groove that accommodates these short
peptides. The peptide binds to the MHC class I molecule in a linear fashion, anchored at
specific positions (usually at the ends of the peptide) within the groove. Example: A viral
peptide might bind within the MHC class I groove, presenting specific amino acid side chains
to the TCR for recognition.
MHC Class II Molecules: These molecules present peptides to CD4+ helper T cells.
The peptides are usually 13-25 amino acids long and are derived from extracellular proteins
processed within the endosomal/lysosomal compartment. The peptide binds in an extended
conformation within the MHC class II binding groove, which is open at both ends, allowing
longer peptides to be accommodated. Example: A bacterial peptide might bind along the
MHC class II groove, exposing key residues to the TCR.
3. Recognition by T Cell Receptors: The T cell receptor (TCR) interacts with the peptide-
MHC complex in a highly specific manner. The TCR recognizes both the peptide and the
MHC molecule, ensuring a precise immune response. The specificity is determined by Peptide
Sequence i.e., specific amino acids in the peptide. Peptide Conformation –the way peptide is
presented within the MHC groove. MHC Type i.e., polymorphic nature of MHC molecules
allows for diverse peptide presentation.

T-cell epitope presented by MHC II

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Functions of T Cell Epitopes:

T cell epitopes are essential components of the immune system, playing a crucial role
in the activation and regulation of T cells.

1. Antigen Recognition

T cell epitopes are specific peptide sequences presented by major histocompatibility

complex (MHC) molecules on the surface of cells. The primary function of these epitopes is
to be recognized by T cell receptors (TCRs) on T cells. This recognition is highly specific,
allowing T cells to distinguish between different antigens and initiate an immune response.

2. Activation of T Cells

The binding of a T cell epitope to a TCR triggers the activation of the T cell. This activation
involves several steps:

✿ Signal Transduction: The engagement of the TCR with the peptide-MHC complex

initiates a signaling cascade within the T cell, leading to its activation.

✿ Clonal Expansion: Activated T cells undergo rapid proliferation, producing a large

number of identical T cells (clones) that can recognize the same epitope.
 ✿ Differentiation: The expanded T cells differentiate into effector T cells (cytotoxic T

cells or helper T cells) and memory T cells.

3. Effector Functions

Activated T cells perform various effector functions depending on their type:

✿ CD8+ Cytotoxic T Cells: These cells recognize epitopes presented by MHC class I

molecules. They directly kill infected or cancerous cells by releasing cytotoxic

molecules like perforin and granzymes, which induce apoptosis in the target cells.
✿ CD4+ Helper T Cells: These cells recognize epitopes presented by MHC class II

molecules. They secrete cytokines that help regulate the immune response by
enhancing the activity of other immune cells, such as B cells, macrophages, and
cytotoxic T cells.

4. Immune Memory

Some activated T cells differentiate into memory T cells, which persist in the body
long-term. These memory T cells provide rapid and robust responses upon re-exposure to the
same antigen. The presence of memory T cells is critical for long-lasting immunity and forms
the basis for the effectiveness of vaccines.

5. Regulation of Immune Responses

T cell epitopes also play a role in regulating immune responses:

★ Cytokine Production: T cells, especially helper T cells, produce various cytokines in

response to epitope recognition. These cytokines can enhance or suppress the activity
of other immune cells, ensuring a balanced immune response.
★ Immunoregulation: Regulatory T cells (a subset of CD4+ T cells) can recognize

specific epitopes and help maintain immune tolerance, preventing autoimmune

reactions.
 Zika virus (ZIKV) infection induces
robust, cross-protective T cell
immunity. In both humans and mice,
ZIKV infection leads to the generation
of Th1 CD4 T cell and effector CD8 T
cell responses, which preferentially
target epitopes in non-structural and
structural proteins, respectively. Studies
have shown that immunity to ZIKV is
cross-protective against subsequent
Dengue virus (DENV) challenge, and
vice-versa. Although studies suggest
CD8 T cells may contribute to
immunopathogenesis in neonatal and
adult mice, with CD4 T cells playing a
potential regulatory role, this remains to
be determined during human infection.
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00820/article_deploy/html/images/cells-08-00820-g001-550.jpg

6. Vaccine Development

The identification and utilization of T cell epitopes are essential for the development
of effective vaccines. Epitope-based vaccines aim to induce strong T cell responses by
including specific peptides that are recognized by T cells. This approach ensures targeted and
efficient immune protection against pathogens.
 T-cell epitope mapping. (1) Immunogens are selected as screen targets; (2) T-cell epitopes are predicted
and tested using various bioinformatic methods; (3) T-cell epitopes are identified by various experimental
methods; (4) Acquisition of candidate epitopes.

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 Workflow for the rational design of T-cell epitope-based vaccines

1. Mapping of T-cell epitopes based on

2. In silico epitope-based vaccine design encompasses various approaches, including structure-based and multi-
epitope-based vaccine design: (a) Epitope-based vaccine design involves fully displaying dominant antigen
epitopes based on protein structure. Design strategies include germline-targeting immunogen, glycosylation
introduction, deletion, stabilization, and epitope grafting. (b) Multi-epitope-based vaccine design integrates the
physicochemical properties of epitopes, arranging them based on their characteristics and utilizing different
linkers to connect epitopes in sequence. (c) Vaccine design in silico leverages various AI technologies.
3. Upon vaccine development and synthesis, reactivity and immunogenicity are analyzed in vitro using ELISA and
Octet.
4. Vaccine candidates identified through the aforementioned steps are subsequently subjected to in vivo immune
assessments to evaluate their efficacy. Evaluation methods include ELIspot, flow cytometry, single-cell RNA
sequencing, Luminex, ELISA, immunohistochemistry, and qPCR.

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7. Therapeutic Applications

Understanding T cell epitopes has led to the development of various therapeutic interventions:

⬥ Cancer Immunotherapy: Tumor-specific T cell epitopes are used to design therapies

that enhance the immune system's ability to target and destroy cancer cells.
⬥ Autoimmune Disease Treatment: Identifying epitopes involved in autoimmune

diseases can help develop therapies that modulate the immune response and reduce
tissue damage.

T-Cell based Cancer Immunotherapy

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 Nanoparticles coated with fragments of the body's own proteins are shown to induce T cells of the immune
system to adopt regulatory functions that suppress autoimmune reactions involving these self-antigens.
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Paratopes:
Paratopes are the antigen-binding sites found on antibodies and T cell receptors
(TCRs). They are crucial for the immune system's ability to recognize and respond to specific
pathogens.

1. Structure and Composition

Paratopes are formed by the variable regions of the heavy and light chains of
antibodies or the alpha and beta chains of TCRs. These variable regions are the most diverse
parts of the molecule, generated through a process called V(D)J recombination.

★ Antibodies: Each antibody has two identical antigen-binding sites (paratopes) located
at the tips of the Y-shaped molecule.
 a) The variable regions of both the heavy (VH) and light (VL) chains contribute
to the formation of the paratope.
b) The complementarity-determining regions (CDRs) within these variable
regions are particularly important for antigen binding. There are three CDRs in
each variable region, making a total of six CDRs per paratope.

Paratopes on the variable regions of the antibodies

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★ T Cell Receptors (TCRs): Each TCR has a single antigen-binding site.

a) The variable regions of the alpha (Vα) and beta (Vβ) chains form the paratope.
b) Similar to antibodies, the CDRs within the variable regions of TCRs are critical
for antigen binding, with three CDRs in each chain.

Why BCR has no paratopes?

BCRs and antigens have direct interactions; interface residues are referred to as
"paratope" on the BCR side and "epitope" on the antigen side. On the other hand, TCRs
engage with peptide fragments originating from antigens, which are displayed by major
histocompatibility complex.
2. Diversity and Specificity

The diversity of paratopes is immense due to the random combination of V, D, and J

gene segments during V(D)J recombination. This process allows the immune system to
generate a vast repertoire of paratopes, each capable of binding a different epitope.

★ Germline Diversity: Different combinations of V, D, and J segments.

★ Junctional Diversity: Addition or deletion of nucleotides at the junctions of V, D, and

J segments.
★ Somatic Hypermutation (for antibodies): Introduces point mutations in the variable

regions of B cells during an immune response, increasing the diversity and affinity of
antibodies.

Diversity in V, D and J segments

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ody-diversity-comes-from-combination-of-variable-and-constant-domain-gene-elements.png

3. Antigen Binding

The paratope's structure is complementary to the epitope on the antigen, allowing for a
highly specific interaction. This binding is driven by various non-covalent forces, such as
hydrogen bonds, electrostatic interactions, Vander Waals forces, and hydrophobic
interactions.

a) Antibodies: The paratope binds to the epitope on the antigen, forming an antigen-
antibody complex. This binding can neutralize the pathogen or mark it for destruction
by other immune cells.
 b) TCRs: The paratope binds to the peptide-MHC complex on the surface of antigen-

presenting cells. This interaction is essential for the activation of T cells and the
subsequent immune response.

Paratope – Antigen binding

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4. Functional Roles

Paratopes play several critical roles in the immune system:

⬥ Antigen Recognition: Paratopes enable antibodies and TCRs to recognize and bind

specific antigens, triggering an immune response.

⬥ Neutralization: By binding to pathogens, antibodies can neutralize their activity and

prevent them from infecting host cells.

⬥ Opsonization: Antibodies bound to antigens facilitate the process of opsonization,

where phagocytic cells (like macrophages and neutrophils) recognize and engulf the
antibody-coated pathogens.
⬥ Complement Activation: The antigen-antibody complex can activate the complement

system, leading to the lysis of the pathogen.

⬥ Cell-Mediated Immunity: TCRs recognizing peptide-MHC complexes on infected or

malignant cells lead to the activation of cytotoxic T cells, which kill these target cells.
 Functions of paratopes
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5. Clinical Applications

Understanding paratopes is essential for developing therapeutic and diagnostic tools:

⬥ Monoclonal Antibodies: These laboratory-produced antibodies are designed to target

specific antigens, using their paratopes to bind with high specificity. They are used in
treating cancers, autoimmune diseases, and infections.
⬥ Vaccines: Knowledge of paratopes and epitopes helps in designing vaccines that elicit

strong and specific immune responses.

⬥ Diagnostic Assays: Paratopes are used in various diagnostic assays, such as ELISAs

and immunohistochemistry, to detect the presence of specific antigens or antibodies.

 HAPTENS
Introduction
Haptens are small molecules, often of low molecular weight, that lack the inherent ability
to provoke an immune response independently. However, they can elicit a robust immune
response when covalently bound to a larger carrier molecule, typically a protein. This duality
underscores their unique role in immunology and their importance in understanding the interplay
between small molecules and the immune system.
The term "hapten" originates from the Greek word "haptein," meaning "to fasten" or "to
bind." The term hapten (half antigen) was introduced in the 1920s by Karl Landsteiner.This
nomenclature aptly describes the fundamental characteristic of haptens: their ability to attach to
larger molecules and, in doing so, form an antigenic complex. While haptens alone are too small
to be recognized by the immune system, their conjugation to a carrier molecule results in a
structure that is large and complex enough to stimulate an adaptive immune response.
The immune response to haptens relies on the recognition of the hapten-carrier conjugate
as a foreign entity by the immune system. This process involves antigen-presenting cells (APCs)
such as dendritic cells, macrophages, and B cells. The carrier protein provides the necessary
structural framework for the immune system to process and present the hapten-carrier conjugate
to T cells, which then activate B cells. The B cells produce antibodies specifically targeting the
hapten moiety, demonstrating the specificity and adaptability of the immune system.

A hapten is an antigen but not an immunogen unless it is attached to a carrier protein. Immunogens are a
subset of antigens that can trigger an immune response on their own.
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Characteristics of Haptens:
Haptens are unique entities with specific characteristics that define their role in
immunology. These traits not only distinguish them from complete antigens but also explain
their behavior and utility in immune responses.
1. Size and Immunogenicity
Haptens are typically small molecules, often with molecular weights below 1,000
Daltons. Their small size and simple structure are insufficient to trigger an immune response on
their own. The immune system requires a certain degree of molecular complexity and size to
recognize a molecule as foreign and initiate an adaptive immune response.
However, when haptens are conjugated to a larger carrier protein, they form a composite
structure known as a hapten-carrier conjugate. This conjugate acts as a complete immunogen,
capable of activating immune cells. The carrier protein provides the necessary bulk and
complexity to ensure recognition by antigen-presenting cells (APCs) and subsequent T-cell
activation, while the hapten acts as a distinct epitope that is specifically targeted by antibodies.
For example, simple chemical groups like dinitrophenyl (DNP) or urushiol (found in
poison ivy) are not immunogenic in isolation. When these haptens bind to proteins in the body,
they become part of a complex that can elicit strong immune responses, often seen as allergic
reactions.
2. Binding Properties
Haptens are chemically reactive molecules capable of forming covalent bonds with
proteins. This property is critical for their role in immune activation. The conjugation process
involves the attachment of haptens to lysine residues or other functional groups on proteins,
forming stable hapten-carrier conjugates.
The covalent nature of the hapten-protein bond ensures that the hapten remains associated
with the carrier molecule during antigen processing. The conjugate is internalized by APCs,
where the protein component is broken down into peptides and presented on the cell surface in
association with major histocompatibility complex (MHC) molecules. Although the hapten itself
is not presented by MHC, its attachment to the carrier molecule ensures its presence in the
immune recognition process, leading to the generation of hapten-specific antibodies.
An example of this property is seen in drug-induced immune reactions. Penicillin, a small
molecule, binds covalently to serum proteins, forming hapten-carrier complexes that can provoke
immune responses.
3. Specificity
One of the most remarkable characteristics of haptens is their specificity. Once the
immune system generates antibodies against a hapten-carrier conjugate, these antibodies
recognize the hapten with high specificity, even in its free, unconjugated form. This reflects the
distinct chemical identity of the hapten, which acts as a unique antigenic determinant (epitope).
This specificity has significant implications in immunology and diagnostics. For instance:
 In allergy testing, synthetic haptens identical to natural allergens are used to detect
specific antibodies in an individual’s serum.
 In research, haptens like DNP or fluorescein are conjugated to antigens or proteins to
study immune responses or track biomolecular interactions using hapten-specific
antibodies.

Antigen presentation on B cells to helper T cells

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Mechanism of Immune Response to Haptens:

The immune response to haptens involves a coordinated interplay between various

components of the immune system. Haptens alone are too small to initiate an immune response,
but when conjugated to a carrier molecule, they become part of a complete antigen that the
immune system can recognize and respond to. The process occurs in distinct stages:
1. Conjugation with Carrier Molecule
The first step in the immune response to haptens is their covalent binding to a carrier
protein, such as albumin, hemocyanin, or synthetic proteins. Haptens are chemically reactive and
often possess functional groups that can form stable covalent bonds with amino acid residues
(commonly lysine) on proteins.

⬥ Purpose of Conjugation: The carrier molecule provides the necessary size and
structural complexity that haptens lack, making the conjugate immunogenic. While
the carrier protein primarily facilitates immune system recognition, the hapten itself
acts as the specific epitope that will be targeted by antibodies.

⬥ Example:
In drug allergies, drugs such as penicillin act as haptens by binding to host proteins in
the bloodstream. The resultant hapten-carrier conjugate becomes the target of an
immune response, leading to hypersensitivity reactions.
2. Antigen Processing and Presentation
Once formed, the hapten-carrier conjugate is taken up by antigen-presenting cells
(APCs), such as macrophages, dendritic cells, or B cells, through a process called endocytosis.
✿ Internalization and Processing: After internalization, the carrier protein portion of the

conjugate is enzymatically degraded within the endolysosomal compartments of the

APC. This breakdown generates peptide fragments derived from the carrier protein,
which are then loaded onto major histocompatibility complex (MHC) class II molecules.
✿ MHC Presentation: The peptide-loaded MHC class II molecules migrate to the surface

of the APC, displaying the processed antigen to T-helper (CD4+) cells. It is important to
note that while the carrier protein is processed and presented, the hapten remains intact
and unprocessed. The presence of the hapten modifies the antigenic properties of the
carrier, effectively creating a unique antigen.
3. Activation of Lymphocytes
The interaction between the APC and T-helper cells marks a critical step in the immune
response. This stage involves two primary cell types:
★ T-helper Cell Activation: T-helper cells recognize the MHC class II-peptide complex

presented on the surface of the APC via their T-cell receptor (TCR). This recognition,
along with co-stimulatory signals provided by the APC, activates the T-helper cells. The
 activated T-helper cells release cytokines that drive subsequent immune processes,
including B-cell activation.
★ B-cell Activation: B cells, which possess surface immunoglobulin (Ig) molecules as

antigen receptors, recognize and bind the hapten-carrier conjugate through the hapten
moiety. The internalized conjugate is processed by the B cell, and carrier-derived
peptides are presented on MHC class II molecules to T-helper cells. This interaction
provides additional activation signals to the B cell, leading to its proliferation and
differentiation into plasma cells.
★ Antibody Production: The plasma cells derived from activated B cells produce

antibodies specific to the hapten. These antibodies are highly specific and can recognize
and bind the hapten even in its free, unconjugated form. This specificity underlines the
immune system's ability to identify and remember molecular patterns.

The Importance of Haptens in Immunology and Their Uses:

Despite their small size and lack of immunogenicity, haptens are crucial to immunology
and have many uses in both medical and scientific domains. Their capacity to create
immunogenic conjugates with bigger molecules has been used to improve diagnostics and
treatments, investigate autoimmune illnesses, create vaccines, and comprehend immune
responses.
1. Allergy Mechanisms
Haptens are central to the understanding of many allergic reactions. Understanding
hapten-mediated allergic mechanisms has informed the development of safer drugs and
desensitization therapies to mitigate allergic responses.

★ Role in Drug Allergies: Small drug molecules like penicillin act as haptens by binding

covalently to host proteins, forming hapten-carrier conjugates. These conjugates are

recognized as foreign by the immune system, triggering hypersensitivity reactions. For
instance, penicillin can bind to serum proteins such as albumin, and the resulting
conjugate may stimulate T-helper cells and B cells, leading to the production of
penicillin-specific antibodies. In sensitized individuals, re-exposure to the drug can cause
severe allergic reactions such as anaphylaxis.
★ Contact Dermatitis: Certain chemicals, such as urushiol from poison ivy or nickel ions,

act as haptens when they bind to skin proteins. The immune system recognizes these
 hapten-protein complexes as antigens, resulting in delayed-type hypersensitivity reactions
and inflammatory responses, characteristic of contact dermatitis.

Hapten induced food allergy mechanism

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M6T8WIK723ZVPA6ZPA1S/hapten+sensitivity.jpg

2. Vaccine Development
Haptens play a pivotal role in the design and development of conjugate vaccines,
especially for pathogens with antigens that are inherently non-immunogenic.
★ Polysaccharide Conjugates: Many bacteria, such as Haemophilus influenzae type b

(Hib), Neisseria meningitidis (meningococcus), and Streptococcus pneumoniae

(pneumococcus), have polysaccharides on their outer surfaces. These polysaccharides are
poorly immunogenic in young children because they do not elicit strong T-cell-dependent
immune responses. By conjugating these polysaccharides to a protein carrier, such as
tetanus or diphtheria toxoid, the immunogenicity is enhanced. The conjugate vaccines
elicit robust T-cell-dependent responses, resulting in the production of high-affinity
antibodies and the establishment of immunological memory.
★ Hapten-Based Therapeutic Vaccines: In cancer immunotherapy, tumor-associated

antigens that act as haptens are conjugated to carriers to enhance immune recognition.
These therapeutic vaccines aim to stimulate the immune system to target and destroy
cancer cells.
 Anti-tumor immunity based on the hapten theory of antibody production
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3. Autoimmunity Research
Haptens serve as valuable tools in understanding autoimmune diseases, where the
immune system mistakenly attacks the body’s own tissues.
★ Hapten-like Behavior of Self-Molecules: In autoimmune diseases, small endogenous

molecules may act as haptens by binding to self-proteins and creating new antigenic
determinants. For example, in drug-induced lupus erythematosus, drugs such as
hydralazine can bind to DNA or nuclear proteins, forming immunogenic complexes that
trigger the production of autoantibodies.
★ Experimental Models: Researchers use haptens like trinitrophenyl (TNP) or
dinitrofluorobenzene (DNFB) to study immune responses in experimental models of
autoimmunity. These models provide insights into the mechanisms by which the immune
system distinguishes between self and non-self, as well as the conditions that lead to the
breakdown of self-tolerance.
 Chemicals and drugs induce autoimmune-like responses via haptenation
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4. Diagnostics and Therapeutics

Haptens have found extensive applications in the fields of diagnostic immunology and
therapeutic research.
⬥ Diagnostic Applications: Haptens are integral to immunoassays, such as enzyme-linked

immunosorbent assays (ELISAs) and radioimmunoassays (RIAs). These techniques use

synthetic haptens to detect specific antibodies or antigens in patient samples. For
example:
 In drug testing, haptens mimicking illicit drugs are used to screen for drug-

specific antibodies.
 In allergy diagnostics, synthetic haptens identical to natural allergens are used to

identify sensitization to specific substances.

⬥ Therapeutic Applications: Synthetic haptens are being explored in targeted therapies.

For instance:
  Haptenization of Tumor Cells: Tumor cells can be modified with haptens to

make them more recognizable by the immune system, promoting anti-tumor

immune responses.
 Hapten-Based Tolerance Induction: In cases of autoimmune diseases or

transplant rejection, haptens are being studied for their potential to induce
immune tolerance by modulating specific immune pathways.
⬥ Drug Design and Development: Understanding hapten-mediated immune reactions has

helped in designing safer pharmaceuticals by modifying chemical structures to minimize

their reactivity as haptens.

ADJUVANTS
Adjuvants are the enhancers of Immune Responses in Immunology. Adjuvants are non-
immunogenic substances that, when administered alongside antigens, significantly enhance the
immune system's response. The term "adjuvant" is derived from the Latin word adjuvare,
meaning "to help," aptly describing their function in immunological applications. These
compounds have transformed the fields of vaccine development and immunotherapy, improving
the efficacy of immunological interventions against infectious diseases, cancers, and
autoimmune disorders.

Role and Importance of Adjuvants in Immunology

Adjuvants are essential components in immunology, ensuring that vaccines and

therapeutic agents effectively elicit strong, long-lasting, and targeted immune responses. Without
the presence of adjuvants, many modern vaccines—particularly those utilizing subunit,
recombinant, or synthetic antigens—would fail to generate sufficient immunity. These antigens,
while safer and more specific than whole-pathogen vaccines, often lack the innate immunogenic
signals required to activate the immune system adequately. Adjuvants overcome this limitation
by enhancing antigen presentation, activating innate immune pathways, and creating a conducive
inflammatory environment.

Key Features of Adjuvants

1. Immune Amplification: Adjuvants significantly boost the magnitude of immune
responses, leading to increased:
 ✿ Antibody Production: Adjuvants stimulate B cells to produce higher quantities

of antibodies, which are essential for neutralizing pathogens.

✿ T-Cell Activation: They enhance the activation of T-helper (CD4+) cells and

cytotoxic T cells (CD8+), which are critical for pathogen clearance and immune
regulation.
✿ Memory Cell Formation: By enhancing the activation and proliferation of

lymphocytes, adjuvants promote the formation of memory B and T cells, ensuring

long-lasting immunity.
2. Directing Immune Responses: Adjuvants play a crucial role in shaping the type of
immune response generated, which is critical for different pathogens and diseases:
✿ Th1 (Cell-Mediated Immunity): Promotes responses against intracellular

pathogens (e.g., viruses, certain bacteria) and cancer cells by activating

macrophages, natural killer (NK) cells, and cytotoxic T cells.
✿ Th2 (Humoral Immunity): Enhances antibody-mediated responses, which are

particularly effective against extracellular pathogens, such as bacteria and

parasites.
✿ This ability to direct the immune response allows for tailored vaccine design,

improving efficacy for specific pathogens or diseases.

3. Dose-Sparing Effect: One of the most significant advantages of adjuvants is their ability
to reduce the quantity of antigen needed per vaccine dose. This "dose-sparing" effect is
particularly beneficial in scenarios such as:
✿ Pandemic Preparedness: During outbreaks, adjuvants enable the production of

more vaccine doses from limited antigen supplies, accelerating vaccination

coverage.
✿ Cost Reduction: By requiring less antigen, adjuvants lower production costs,

making vaccines more accessible, especially in resource-limited settings.

4. Enhanced Immunological Memory: Adjuvants promote the development of long-term
immunological memory by stimulating the differentiation and survival of memory B and
T cells. This ensures that the immune system can respond more rapidly and effectively
upon subsequent exposure to the pathogen.
5. Broadened Antigen Recognition: By enhancing antigen presentation and increasing the
diversity of peptides displayed on major histocompatibility complex (MHC) molecules,
 adjuvants improve the immune system's ability to recognize and respond to a wider range
of pathogen variants.
6. Improved Responses in Vulnerable Populations
✿ Elderly Individuals: Aging often leads to a decline in immune function.

Adjuvants can boost vaccine efficacy in older adults, who are more susceptible to
infections.
✿ Infants and Immunocompromised Individuals: In these groups, where immune

responses are naturally weaker, adjuvants help generate protective immunity.

7. Reduction in Booster Doses: By amplifying the initial immune response, adjuvants can
reduce the number of booster doses required, simplifying vaccination schedules and
improving compliance.
Examples, highlighting the Role of Adjuvants
1. Hepatitis B Vaccine: The hepatitis B vaccine uses aluminum salts (alum) to enhance the
immune response to the recombinant surface antigen. Without alum, the vaccine would
be significantly less effective at generating protective immunity.
2. COVID-19 Vaccines
 The Novavax vaccine uses the Matrix-M adjuvant, which enhances both humoral

and cell-mediated immunity, ensuring robust protection against SARS-CoV-2.

 Adjuvants like AS03 and CpG 1018 have been employed in other COVID-19

vaccine candidates to improve immunogenicity and dose efficiency.

3. Cancer Vaccines: In cancer immunotherapy, adjuvants such as TLR agonists (e.g.,
monophosphoryl lipid A) are used to stimulate strong anti-tumor immune responses by
activating dendritic cells and promoting cytotoxic T-cell activity.

Adjuvants' various functions in enhancing the body's immunological responses

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Mechanisms of Action of Adjuvants

Adjuvants employ a range of mechanisms to enhance, direct, and sustain immune

responses. By interacting with various components of the immune system, they amplify antigen
recognition, recruit immune cells, and promote the development of adaptive immunity. Some
important mechanisms:
1. Prolonged Antigen Availability: Many adjuvants, such as aluminum salts (alum), form a
physical depot at the site of injection. This depot gradually releases the antigen over time,
ensuring prolonged stimulation of the immune system, maintains antigen availability, allowing
for repeated stimulation of antigen-presenting cells (APCs), and enhances the chance of adaptive
immune cells encountering the antigen.
2. Activation of Innate Immunity with Pattern Recognition Receptors (PRRs): Adjuvants
interact with PRRs on APCs, such as Toll-like receptors (TLRs) and NOD-like receptors
(NLRs). These receptors detect pathogen-associated molecular patterns (PAMPs) or damage-
associated molecular patterns (DAMPs) introduced or mimicked by adjuvants. It strengthens the
"danger signal" needed to initiate a robust immune response.

Example Pathways:
a) TLR agonists (e.g., monophosphoryl lipid A) stimulate TLR4, leading to
the activation of nuclear factor kappa-light-chain-enhancer of activated B
cells (NF-κB).
b) This induces the production of pro-inflammatory cytokines (e.g.,

interleukin-1 (IL-1), IL-6, tumor necrosis factor-alpha (TNF-α)), which

recruit and activate other immune cells.
3. Enhanced Antigen Uptake - Adjuvants promote the efficient uptake of antigens by APCs,
such as dendritic cells and macrophages. Antigens are processed into peptides and presented on
MHC class I (for cytotoxic T-cell activation) or MHC class II (for helper T-cell activation)
molecules. This process involves:
 Phagocytosis: Internalization of particulate antigens.

 Pinocytosis: Uptake of soluble antigens.

4. Stimulation of Cytokine Secretion: Adjuvants induce the release of cytokines and
chemokines, which amplify and guide the immune response:
 Th1 Response: Promoted by cytokines like interferon-gamma (IFN-γ) and

interleukin-12 (IL-12), leading to cell-mediated immunity.

 Th2 Response: Enhanced by cytokines like interleukin-4 (IL-4) and IL-10,

fostering antibody production.

Example: synthetic DNA sequences mimicking bacterial DNA, stimulate TLR9
to trigger a Th1-biased response.
5. Generation of Immune Responses
Adjuvants can influence the type of immune response based on their composition and mode of
action:
⬥ Th1 Response:

★ Targets intracellular pathogens (e.g., viruses, intracellular bacteria).

★ Promoted by adjuvants like TLR agonists, which stimulate IFN-γ and activate

cytotoxic T cells and macrophages.

Example: The AS01 adjuvant system in malaria vaccines enhances Th1-biased
immunity.
⬥ Th2 Response:

★ Effective against extracellular pathogens (e.g., bacteria, parasites).

★ Induced by adjuvants like alum, which promote IL-4 secretion and antibody

production.
Example: Alum-based vaccines for diphtheria and tetanus rely on Th2 responses
to neutralize toxins.
 Mechanisms of action of adjuvants
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Different Types of Adjuvants in Immunology:

Adjuvants are crucial substances that enhance the body's immune response to an antigen.
Without adjuvants, many vaccines—particularly those derived from subunit, recombinant, or
synthetic antigens—would not produce an immune response of sufficient strength. These
compounds are vital in modern immunology for boosting vaccine effectiveness, promoting long-
term immunity, and enabling targeted treatments. Adjuvants are categorized based on their
composition, function, and the way they enhance the immune system's response.

1. Inorganic Adjuvants
Inorganic adjuvants, composed mostly of salts and minerals, are commonly employed in
vaccines due to their ability to promote immune activation through depot formation or by
stimulating the innate immune system.
 ★ Aluminum Salts (Alum): The most frequently used adjuvants, aluminum salts like

aluminum hydroxide, aluminum phosphate, and aluminum sulfate, create a depot at the
injection site. This allows for slow release of the antigen, providing prolonged exposure
and enhancing immune activation. These adjuvants are particularly effective in triggering
Th2-type immune responses, resulting in high antibody production. Applications:
Diphtheria, tetanus, hepatitis B, and Hib vaccines.
★ Calcium Phosphate: Similar to aluminum salts, calcium phosphate forms a depot that

provides extended antigen release, ensuring continuous immune stimulation.

Applications: Hepatitis B and various experimental vaccines.
2. Oil-in-Water Emulsions
Oil-in-water emulsions are adjuvants made by combining oil droplets with water to form
a suspension. These emulsions are known for enhancing both humoral (antibody-mediated) and
cellular (T-cell mediated) immunity.
★ MF59 is an oil-in-water emulsion that contains squalene (a natural oil) mixed with water

and surfactants, improves antigen uptake by antigen-presenting cells (APCs). It helps

promote a balanced Th1/Th2 immune response. Applications: Influenza vaccines,
especially for elderly individuals, as it increases the strength and duration of immune
protection.
★ AS03 is another oil-in-water emulsion that also includes squalene and surfactants. It

helps stimulate both innate and adaptive immune responses and is noted for improving
immune responses even with lower antigen doses. Applications: H1N1 influenza
vaccines and other pandemic flu vaccines.
3. Lipid-Based Adjuvants
These adjuvants are made from lipid particles that enhance immune cell activation,
particularly in stimulating cellular immunity.
★ ISCOMs (Immunostimulating Complexes): ISCOMs are lipid complexes formed from

cholesterol and other lipids, which encapsulate antigens and serve as a delivery system to
present the antigen to immune cells. They are highly effective in inducing both cellular
and humoral immune responses. Applications: HIV, malaria, and experimental cancer
vaccines.
★ QS-21 is a potent saponin-based adjuvant derived from the Chilean soapbark tree. It

activates both innate and adaptive immune responses, often inducing Th1 responses and
boosting antibody production. It is commonly combined with other adjuvants to enhance
 their effectiveness. Applications: Cancer vaccines and malaria vaccines (under clinical
investigation).
4. Toll-Like Receptor (TLR) Agonists:
TLRs are critical components of the innate immune system, responsible for detecting
pathogen-associated molecular patterns (PAMPs). TLR agonists act as adjuvants by activating
TLRs and initiating immune cascades that enhance the immune response.
★ Monophosphoryl Lipid A (MPL): MPL is a derivative of lipopolysaccharide (LPS)

from bacteria, which stimulates TLR4 and activates a robust Th1 immune response,
enhancing cell-mediated immunity. Applications: Cervarix (HPV vaccine), malaria
vaccines, and cancer immunotherapies.
★ CpG Oligodeoxynucleotides (CpG ODN): CpG ODN are synthetic DNA sequences

that resemble bacterial DNA, recognized by TLR9. They stimulate a Th1 immune
response and promote cytokine production, such as interferon-gamma (IFN-γ).
Applications: Cancer vaccines and certain antiviral vaccines.
5. Saponin-Based Adjuvants:
Saponins are natural plant-derived compounds that can activate both humoral and cellular
immune responses. They are particularly effective in activating dendritic cells, which are crucial
for initiating immune reactions.
★ QS-21 (as mentioned earlier): QS-21 is a well-known saponin-based adjuvant that

boosts the immune system’s ability to recognize and process antigens, often used
alongside other adjuvants to enhance the overall immune response.
★ ISCOMs (Immunostimulating Complexes): ISCOMs include saponins along with

lipids, helping the immune system recognize and process antigens more effectively.
6. Bacterial-Derived Adjuvants
Some bacterial components also serve as adjuvants by activating immune responses and
bolstering the body’s defense mechanisms.
★ Bordetella pertussis toxin: This toxin, derived from the bacteria responsible for

whooping cough, increases the immunogenicity of vaccines by stimulating the immune

system. Applications: DTP (Diphtheria, Tetanus, Pertussis) vaccines.
★ Heat-Labile Toxin (LT): LT, derived from Escherichia coli, promotes both cellular and

humoral immune responses by activating innate immune cells. Applications: Used in

various vaccines to boost immune responses.
7. Other Adjuvants
★ Carbohydrate-Based Adjuvants: Derived from bacterial polysaccharides, these
adjuvants are particularly useful in vaccines targeting bacterial infections. When
conjugated with proteins, they enhance the immune response, especially against
encapsulated bacteria.
Examples: Polysaccharide-protein conjugates in vaccines for Haemophilus
influenzae type b (Hib), pneumococcus, and meningococcus.

Types of adjuvants and their roles

Image Source: https://www.mdpi.com/2076-393X/11/2/453

Applications of Adjuvants in Immunology:

Adjuvants play an essential role in modern immunology, significantly enhancing immune
responses to antigens. By boosting the body’s immune system, adjuvants are integral in a variety
of applications, ranging from vaccine development to therapeutic treatments. Their ability to
amplify and direct immune responses makes them indispensable in both preventative and
therapeutic approaches.
1. Vaccine Development
Adjuvants are most commonly used in vaccines to improve their effectiveness. Vaccines
that rely on subunit, recombinant, or synthetic antigens often require adjuvants to generate a
strong and lasting immune response. Without adjuvants, these vaccines may not trigger sufficient
immunity.
✿ Enhancing Immune Responses: Adjuvants increase antibody production and stimulate

T-cell activation, leading to stronger and more robust immunity. This is particularly
important for vaccines where both humoral (antibody-mediated) and cellular (T-cell-
mediated) immunity are needed.
✿ Reducing Antigen Dosage: By boosting immune responses, adjuvants allow for lower

doses of the antigen in vaccines. This can help stretch limited vaccine supplies, which is
especially useful during outbreaks or pandemics when rapid vaccine distribution is
necessary.
✿ Promoting Long-Term Immunity: Adjuvants enhance the generation of memory B-

cells and T-cells, providing longer-lasting protection and preventing reinfection,

particularly in diseases prone to mutation, such as influenza or COVID-19.
Examples in Vaccine Applications:
a) Aluminum Salts (Alum): Commonly used in vaccines like diphtheria, tetanus, hepatitis
B, and Hib.
b) Oil-in-Water Emulsions (e.g., MF59): Used in influenza vaccines, particularly for older

populations.
c) Toll-Like Receptor (TLR) Agonists (e.g., MPL): Used in HPV and malaria vaccines.

Aluminium salts as vaccine adjuvants

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2. Cancer Immunotherapy
Adjuvants are utilized in cancer immunotherapy to activate the immune system, enabling
it to target and attack tumor cells more effectively.
✿ Enhancing Tumor Antigen Recognition: Adjuvants can boost the immune system's

ability to identify tumor-associated antigens (TAAs), improving the targeting and

destruction of cancer cells.
✿ Stimulating T-cell Activation: Certain adjuvants activate cytotoxic T lymphocytes

(CTLs), which are essential for destroying cancer cells.

✿ Improving Cancer Vaccine Effectiveness: When combined with cancer vaccines,

adjuvants enhance the immune response against specific cancer antigens, improving
vaccine efficacy.
Examples in Cancer Immunotherapy:
a) ISCOMs (Immunostimulating Complexes): Used in experimental cancer vaccines.
b) QS-21: Combined with other adjuvants to enhance cancer vaccine responses.

The immunogenicity of vaccinations is improved by adjuvants. A T helper-polarizing cytokines, antibodies, and activated
T cells are produced in small amounts by vaccines devoid of adjuvants. B On the other hand, vaccines that contain
adjuvants encourage the maturation of more APCs, enhance the interaction between APCs and T cells, and encourage the
production of more T helper-polarizing cytokines, multifunctional T cells, and antibodies. These factors result in broad
and long-lasting immunity as well as dose and antigen savings. This figure was created with BioRender
(https://biorender.com/)
Image source: https://media.springernature.com/m685/springer-static/image/art%3A10.1038%2Fs41392-023-01557-
7/MediaObjects/41392_2023_1557_Fig1_HTML.png
3. Autoimmune Disease Research
Adjuvants are valuable tools in studying autoimmune diseases, providing insights into
how the immune system mistakenly attacks the body’s own tissues. By inducing inflammation
and immune activation, adjuvants help researchers explore the underlying mechanisms of
autoimmune responses.
✿ Autoantigen Presentation: Some adjuvants stimulate the immune system to produce

autoantibodies that attack self-molecules, aiding in the study of autoimmune conditions

like rheumatoid arthritis and multiple sclerosis.
✿ Modulating Immune Responses: In autoimmune diseases, specific adjuvants can help

adjust immune responses to reduce attacks on healthy tissues, contributing to the

development of treatments that restore immune tolerance.
Example:
a) Bacterial-derived adjuvants like Bordetella pertussis toxin: Used in animal models to
study autoimmune mechanisms.
4. Allergy and Hypersensitivity Treatment
Adjuvants are used in allergy treatment to modify the immune system's response to
allergens. They are incorporated into allergy vaccines (allergen immunotherapy) to help
desensitize the immune system to specific allergens.
✿ Modulating Immune Responses: Adjuvants can shift immune responses from a Th2-

dominant response (associated with allergies) to a more balanced Th1/Th2 response,

reducing hypersensitivity.
✿ Enhancing Allergen-Specific Immunity: By boosting immune tolerance to allergens,

adjuvants provide long-term relief for individuals with allergic conditions like hay fever
or asthma.
Example:
a) Aluminum salts and oil-in-water emulsions: Used in vaccines for pollen or dust mite
allergies.
5. Infectious Disease Management
Adjuvants play an essential role in enhancing the immune system's ability to combat
infectious diseases. They are used to improve the efficacy of vaccines against a wide variety of
pathogens, including viruses and bacteria.
 ✿ Broadening Immune Responses: Adjuvants enhance both antibody production and

cellular immunity, enabling the immune system to target a broader range of pathogens
and improving vaccine efficacy.
✿ Improving Vaccine Efficacy in Special Populations: Adjuvants are particularly

important for vaccines targeting vulnerable groups, such as infants, the elderly, and
immunocompromised individuals, who may have weaker immune responses to vaccines.
Examples in Infectious Disease Vaccines:
a) MF59 and AS03: Used in influenza vaccines for at-risk populations.
b) MPL and CpG ODN: Used in vaccines for malaria, tuberculosis, and hepatitis.

6. Therapeutic Applications in Chronic Diseases

Adjuvants are also being explored for use in the treatment of chronic diseases, such as
chronic viral infections (e.g., HIV) and autoimmune disorders, where modulation of the immune
response is necessary.
✿ Chronic Viral Infections: Adjuvants help boost immune responses against persistent

viral infections, making them valuable in the development and treatment of HIV
vaccines.
✿ Autoimmune Disease Therapy: In autoimmune conditions, adjuvants can help adjust

immune responses to prevent the immune system from attacking healthy tissues.
Example:
a) ISCOMs and QS-21: Investigated in HIV and chronic viral infections.
7. Diagnostics
Adjuvants are used in diagnostic tools to improve the detection of specific antibodies or
antigens, enhancing the sensitivity of tests such as enzyme-linked immunosorbent assays
(ELISA) and lateral flow assays.
✿ Boosting Sensitivity: Adjuvants enhance the immune response to antigens or antibodies,

increasing the accuracy and reliability of diagnostic tests.

Examples:
Saponins and TLR agonists: Used in diagnostic applications for detecting specific
infectious agents or antibodies.
 Factors influencing Imunogenecity

The ability of a material, usually a foreign molecule (antigen), to trigger an immune

response is known as immunogenicity, and it is a crucial idea in immunology. Innate and
adaptive immune system activation may be a part of this reaction, which can lead to antibody
formation, T cell activation, and the development of immunological memory. The two main
kinds of factors affecting immunogenicity are host-related factors and antigen-related factors.
The duration, specificity, and intensity of the immune response are all influenced by these
variables.
Factors Associated with Antigens:
1 Size of the Antigen: The size of molecules Compared to smaller molecules, larger ones
are more immunogenic. This is due to the fact that immune cells are better able to
recognise and handle bigger molecules. High molecular weight proteins and
polysaccharides frequently elicit more robust immune responses. Conversely, for tiny
molecules (haptens) to elicit an immune response, they might need to conjugate with a
carrier protein.

2 Complexity of Molecular Structure: Proteins, lipoproteins, and glycoproteins are

examples of complex molecules with a variety of shapes that are highly immunogenic
because they offer several epitopes for immune recognition. Simpler molecules, such as
carbohydrates or lipids, are less immunogenic because they contain fewer or less diverse
epitopes.

3 Foreignness: The immunogenicity of an antigen is influenced by how much the immune

system views it as "non-self". An immune response is more likely to be triggered by
antigens that are more different from the host's own molecules. Self-tolerance
mechanisms frequently stop the immune system from reacting to the proteins in the host.

4 The stability and chemical composition: For recognition and processing, an antigen's
stability and integrity are essential. Long-lasting compounds that are highly stable can
 interact with immune cells in an efficient manner. Antigens that are denatured or
degraded may no longer have the capacity to activate the immune system.

5 Form Physical: In general, antigens in aggregated or particulate form elicit stronger

immune responses than those in soluble form. Immune activation is improved by antigen
presenting cells' (APCs) increased uptake of particulate antigens.

6 The dose of antigens: The immune response is affected by the quantity of antigen that is
injected into the body. An overly high dose can cause immunological tolerance or
suppression, while a dose that is too low may not be able to activate immune cells.
Frequently, there is a dose that maximises immunogenicity.

7 Administrative Route: The kind and intensity of the immune response are influenced by
the way an antigen is presented. As an example, Strong systemic reactions are often
elicited by subcutaneous and intramuscular methods. Weaker reactions or tolerance may
be brought on by intravenous methods. Localised immunological responses in mucosal
tissues can be triggered by mucosal channels, such as the nasal or oral.

8 Presence of Adjuvants: Adjuvants are chemicals that are included in vaccinations to

improve the immune system's reaction to an antigen. They work by attracting immune
cells to the site of antigen presentation, enhancing innate immunity, or extending antigen
exposure. Toll-like receptor (TLR) agonists, squalene-based emulsions, and aluminium
salts are a few examples.

Host-Related Factors Influencing Immunogenicity:

1. Genetic Background
The genetic makeup of an individual plays a pivotal role in shaping the immune response
to antigens. Key elements include:
★ Major Histocompatibility Complex (MHC): The diversity and types of MHC

molecules (known as Human Leukocyte Antigens, or HLA, in humans) influence how

antigens are processed and presented to T cells. MHC class I molecules present
 endogenous antigens (e.g., from viruses) to CD8+ cytotoxic T cells, while MHC class II
molecules present exogenous antigens to CD4+ helper T cells. Genetic polymorphisms in
MHC genes determine the range of peptides that can be effectively presented, affecting
immunogenicity.
★ Variability in Immune Receptor Genes: Differences in genes encoding immune

receptors like Toll-like receptors (TLRs) and immunoglobulin genes can impact
recognition and response to antigens. For example, certain alleles may confer
susceptibility or resistance to infections or influence vaccine efficacy.
2. Age
Age significantly impacts the efficiency and nature of immune responses:
★ Neonates (Newborns): Neonatal immune systems are immature, with limited antigen-

presenting cell function and reduced production of pro-inflammatory cytokines. They

rely heavily on maternal antibodies transferred through the placenta or breast milk for
immunity. This immaturity results in weaker responses to vaccines and infections.
★ Elderly Individuals: Aging leads to immunosenescence, characterized by:

 Reduced production of naive T and B cells due to thymic involution and bone

marrow changes.
 A shift towards a memory-biased immune repertoire, which may result in

suboptimal responses to novel antigens.

 Impaired innate immunity, such as decreased phagocytosis and reduced dendritic

cell function. These changes contribute to reduced vaccine efficacy and increased
susceptibility to infections.
3. Health Status
The overall health of an individual profoundly influences immunogenicity:
★ Immunocompromised States: Conditions such as HIV/AIDS, cancer, malnutrition, or

the use of immunosuppressive therapies (e.g., chemotherapy, corticosteroids) impair

immune cell function. This results in weakened responses to antigens, including vaccines
and infections.
★ Autoimmune Diseases: In autoimmune conditions, the immune system may exhibit

heightened sensitivity or dysregulation, leading to inappropriate responses to self-

antigens. This hyper-responsiveness can also influence reactions to foreign antigens.
 ★ Chronic Diseases: Chronic conditions like diabetes or cardiovascular disease can impair

immune system function, potentially reducing immunogenicity.

4. Previous Exposure
The immune system’s ability to "remember" antigens through immunological memory plays a
vital role:
★ Primary Exposure: The first encounter with an antigen generates a relatively slow and

less robust immune response, characterized by the activation of naive T and B cells.
★ Secondary Exposure: Subsequent encounters with the same or cross-reactive antigens

trigger memory B and T cells, resulting in faster, stronger, and more specific immune
responses. This principle underlies the effectiveness of booster vaccines.
★ Cross-Reactivity: Previous exposure to structurally similar antigens can result in cross-

reactivity, where memory cells react to related antigens.

5. Hormonal Factors
Hormones have a regulatory effect on the immune system, influencing immunogenicity:

★ Corticosteroids: These hormones, released during stress or administered therapeutically,

suppress immune responses by reducing inflammation and inhibiting T cell activation.

★ Sex Hormones:
 Estrogens (predominantly in females) generally enhance immune responses,
contributing to the higher prevalence of autoimmune diseases in women.
 Androgens (predominantly in males) tend to suppress immune responses, which
may contribute to differences in disease susceptibility and vaccine efficacy
between sexes.
★ Pregnancy: During pregnancy, the immune system adapts to tolerate the fetus (a semi-
allograft) by reducing T cell activity and pro-inflammatory responses. This can decrease
immunogenicity to certain infections or vaccines during this period.
6. Microbiome: The microbiome, the collection of microorganisms living in and on the human
body, profoundly influences immune system function:
★ Immune System Modulation: A diverse and balanced microbiome helps educate and

regulate the immune system, promoting effective responses to antigens while preventing
overreactions (e.g., allergies or autoimmune diseases).
 ★ Dysbiosis: Disruption of the microbiome balance (e.g., due to antibiotics, diet, or

infections) can impair immune responses or lead to excessive inflammation.

★ Microbial Adjuvants: Commensal microbes can act as natural adjuvants, enhancing
the immunogenicity of antigens by stimulating innate immune pathways.
Environmental Factors Influencing Immunogenicity:
Environmental factors play a significant role in shaping the immune system’s ability to
respond to antigens. These factors interact with both antigen- and host-related elements,
influencing the strength, type, and duration of immune responses.
1. Exposure to Pathogens
The frequency and diversity of pathogen exposure significantly impact the immune
system's readiness and capacity to respond to new antigens:
★ Immune Priming: Repeated exposure to pathogens or their antigens trains the
immune system to recognize and respond more efficiently to future challenges. For
example:
 Early childhood exposure to various infections can enhance the development
of a robust immune system, often referred to as the "hygiene hypothesis."
 Exposure to cross-reactive antigens may result in partial immunity to related
pathogens (e.g., cross-reactivity between influenza virus strains).

★ Pathogen Burden: High levels of chronic exposure to pathogens, especially in

regions with poor sanitation or endemic diseases, can overburden the immune system,
reducing its ability to respond effectively to vaccines or new infections.

★ Vaccination History: Vaccination simulates pathogen exposure under controlled

conditions, effectively priming the immune system without causing disease. This
improves immunogenicity by inducing memory B and T cell formation.
★ Global Variation: Differences in pathogen exposure due to geographical location
(e.g., tropical vs. temperate climates) can lead to population-specific variations in
immune system development and vaccine responses.
2. Nutrition
Nutrition is a cornerstone of immune health, directly influencing immunogenicity
through the availability of nutrients essential for immune cell function and antigen recognition.
 ★ Micronutrient Deficiencies: Deficiencies in key vitamins and minerals impair
immune responses and reduce immunogenicity:
 Vitamin A: Supports epithelial barriers, promotes T cell function, and
enhances antibody production.
 Vitamin D: Regulates innate and adaptive immune responses, particularly
through its effects on macrophages, dendritic cells, and T cells.
 Vitamin C: Enhances neutrophil function, supports oxidative burst activity,
and stabilizes immune cell membranes.
 Vitamin E: Acts as an antioxidant, protecting immune cells from oxidative
stress and supporting T cell function.
 Zinc: Critical for thymic function, T cell development, and cytokine
production.
 Iron: Essential for the proliferation and function of immune cells, including
lymphocytes.
★ Malnutrition: Protein-energy malnutrition weakens both innate and adaptive
immunity by reducing the production of immune cells, cytokines, and antibodies.

★ Overnutrition and Obesity: While undernutrition suppresses immunity,

overnutrition and obesity are associated with chronic low-grade inflammation, which
can impair vaccine responses and lead to immune dysfunction.
★ Diet Composition: Diets rich in antioxidants, omega-3 fatty acids, and probiotics can
enhance immune function, while diets high in refined sugars and unhealthy fats may
suppress immune responses.
3. Stress and Lifestyle
Lifestyle factors, including stress, sleep patterns, and habits, play a significant role in
determining immunogenicity.
★ Chronic Stress: Prolonged stress activates the hypothalamic-pituitary-adrenal (HPA)
axis, leading to the release of glucocorticoids (e.g., cortisol), which suppress immune
functions:
 Reduced cytokine production.
 Impaired activity of antigen-presenting cells (APCs) and T cells.
  Lower antibody responses to vaccines.
★ Poor Sleep: Sleep is critical for immune regulation. During deep sleep, cytokine
production increases, supporting immune memory formation. Sleep deprivation
disrupts this process, leading to:
 Reduced vaccine efficacy.
 Lower T cell activation and antibody production.
 Increased susceptibility to infections.

★ Unhealthy Habits:
 Smoking: Smoking damages epithelial barriers, increases oxidative stress,
and impairs the function of innate and adaptive immune cells, reducing overall
immunogenicity.
 Alcohol Consumption: Excessive alcohol intake suppresses the immune
system by reducing cytokine production, impairing macrophage and T cell
functions, and depleting essential nutrients.
 Physical Inactivity: Sedentary lifestyles are linked to inflammation and
reduced immune competence, while moderate regular exercise enhances
immunogenicity by improving circulation of immune cells and reducing
inflammation.
★ Environmental Pollutants: Exposure to air pollution, heavy metals, and industrial
chemicals can cause oxidative stress and inflammation, impairing immune responses
and reducing vaccine efficacy.
 STRUCTURE OF ANTIBODIES
Antibodies, also called immunoglobulins, are protein molecules produced naturally by B-
lymphocytes. The term "antibody" refers to an immunoglobulin that specifically recognizes and
binds to an epitope on antigens. These soluble molecules, secreted by plasma cells, circulate
throughout the body to locate and attach to foreign substances, or antigens. When antibodies bind
to an antigen or microbial epitope, they can inhibit microbial activity and spread through various
mechanisms, such as immobilization, preventing microbial attachment to host cells, enhancing
phagocytosis, or marking microbes for destruction by soluble molecules or immune cells like
natural killer (NK) cells and eosinophils.
The primary role of antibodies is to recognize antigens and bind to them at their specific
antigen-binding sites. Once an antigen is detected, B-lymphocytes undergo proliferation and
differentiation into plasma cells, which then produce large quantities of antibodies to combat the
antigen. Their unique structure ensures antigen specificity, allowing them to bind exclusively to
their corresponding antigens. There are two classes of these antibodies:
1 Membrane bound antibodies: B-cell receptors (BCR) are composed of membrane-
bound antibodies, sometimes referred to as membrane-bound immunoglobulin (mIg) or
surface immunoglobulins (sIg). The surface of antibody-bound B-cells has receptors that
enable the B cell to recognise a particular antigen in the body. This activation causes the
B cell to multiply and develop into plasma cells, which generate the circulating
antibodies. The B-cell receptor (BCR) is typically made up of IgD or IgM antibodies that
are surface-bound and linked to Ig-α and Ig-β heterodimers, which can trigger signal
transduction. Between 50,000 and 100,000 antibodies are attached to the surface of a
single B-cell.
2 Freely circulating antibodies: The effector mechanisms of humoral immunity are
provided by freely circulating antibodies in the blood, which either hunt for and
neutralise antigens or mark them for removal by immune cells such as antigen-presenting
cells.
Structural Details:
Antibodies are large, globular, plasma proteins that weigh around 150 kDa and have a
diameter of 10 nm. Antibodies are glycoproteins because they are composed of glycans, which
are sugar chains that are attached to conserved amino acid residues. Glycans are essential for
maintaining the structure and functionality of antibodies. Additionally, they change the
antibody's affinity for the FcR (s). Immunoglobulin (Ig), the fundamental functional unit of an
antibody, is monomeric, whereas the released antibodies might be dimeric, tetrameric,
pentameric, or polymeric. The antibody is composed of constant C regions and variable V
regions.

Using myeloma proteins, Edelman for the first time extracted the antibody from a
multiple myeloma blood sample and initially identified its structure. The molecular weights of
the two chains they identified were 20 kDa for the light chain and 50 kDa for the heavy chain.
According to their respective concentrations, the fundamental antibody unit was made up of two
heavy and two light chains. Thus, the Antibodies are heterodimers. In IgG, IgD, and IgE, this
four-chain structure is present. In contrast, IgM exists as a pentamer with five basic units, and
IgA exists in both monomeric and polymeric forms (consisting of multiple basic four-chain unit
structures).

Immunoglobulin domains
 The Immunoglobulin monomer has a “Y” – shaped molecule which has 4-polypeptide

chains; two identical heavy (H) chains and two identical light (L) chains which are
connected by a disulfide bond.
 Each of the chains is made up of globular domains known as immunoglobulin domains

formed by intra-chain disulfide bonds. The domains containing 70-110 amino acids and
are classified into different types depending on size and function such as the variable
domain (IgV) and the constant domains (IgC).
 The single variable region or domains (IgV) is of two forms, VL and VH consisting of 100

amino acids and the constant domains (CH and CL).

 The domains have a specific immunoglobulin fold which has two beta-sheets sheets form

a sandwich shape, that is held together by the forces between the interaction of conserved
cysteines and other amino acids.
 The domains have a particular immunoglobulin fold with two beta-sheets that form a

sandwich shape. The interaction of conserved cysteines and other amino acids holds the
sheets together.
 General structure of Antibody
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The Heavy Chain of Antibodies:

In mammals, immunoglobulin heavy chains exist in five types, identified by the Greek
letters α, δ, ε, γ, and μ. These heavy chains determine the class of immunoglobulin, meaning
each immunoglobulin isotype contains a specific type of heavy chain (e.g., μ chains in IgM and γ
chains in IgG). The heavy chain's structural differences contribute to the antibody's function and
flexibility, allowing it to bind specifically to antigens and perform its immune role effectively.
The heavy chains vary in size and composition:
 α and γ chains consist of approximately 450 amino acids.
 μ and ε chains are slightly larger, containing about 550 amino acids.
Each heavy chain is divided into two regions:
1. Constant (C) Region:
⬥ Identical across all antibodies of the same isotype but different between

antibodies of different isotypes.

⬥ For γ, α, and δ chains, the constant region contains three immunoglobulin

domains and a hinge region that provides flexibility to the antibody.

 ⬥ For μ and ε chains, the constant region has four immunoglobulin domains but

lacks a hinge region.

2. Variable (V) Region:

⬥ Unique to antibodies produced by different B cells but identical in antibodies

derived from the same B-cell clone.

⬥ Approximately 110 amino acids long and composed of a single immunoglobulin

domain.

The Light Chain of Antibodies:

By working in concert with the heavy chains, the light chains enhance the antibody's
ability to recognize and bind specific antigens, playing a critical role in immune defense. In
mammals, antibodies contain light chains that are of two types: lambda (λ) and kappa (κ). Each
light chain consists of two domains:
1. Variable (V) Domain: Responsible for antigen binding and varies among antibodies
produced by different B cells.
2. Constant (C) Domain: Conserved within a particular type of light chain (λ or κ) and
contributes to the structural stability of the antibody.

Important Features of the Light Chain:

★ The light chain is composed of 211 to 217 amino acids in length.

★ Each antibody molecule contains two identical light chains of the same type (either λ or

κ).
★ Only one type of light chain (either λ or κ) is present per antibody, regardless of the

heavy chain isotype. For example, an IgG antibody may pair its heavy chains with either
two λ chains or two κ chains but never a mix of both.
★ Light chains pair with heavy chains to form the antigen-binding sites of the antibody.

Together, the variable domains of the heavy and light chains create the specific structure
that recognizes and binds to antigens.
★ The light chain does not determine the antibody isotype but contributes to the diversity

and specificity of antigen binding.

★ The heavy and light chains are antigenically distinct, meaning they are recognized as

separate molecular entities by the immune system.

 ★ The pairing of a single type of light chain (λ or κ) with the heavy chains ensures

structural consistency within each antibody molecule.

Heavy and light chains of an antibody

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Antibody Hypervariable Regions:

An antibody's hypervariable regions are essential structural components that give it the
capacity to identify and bind particular antigens. These areas make up the heart of the antigen-
binding site, commonly referred to as the paratope, and are found in the variable domains of both
the heavy and light chains. The hypervariable regions collectively form the antigen-binding site
of the antibody, where specific interactions with the antigen occur.
There are three hypervariable areas in each of the heavy (VH) and light (VL) chains'
variable domains, for a total of six hypervariable regions in an antibody's antigen-binding site.
More conserved sections known as framework regions (FRs) encircle these areas, provide
structural support and preserving the variable domain's general form.
Because they establish the complementarity between the antibody and its target antigen, the
hypervariable sections are also known as complementarity-determining regions (CDRs).
The Cluster of Differentiation, Fv, Fab, and Fc Regions of Antibodies:
Antibodies are composed of distinct structural regions, each contributing to their function
in antigen recognition and immune response. These regions include the Fab, Fv, and Fc, which
collectively define the antibody's role in antigenicity and immune activation.
1. Fab Region (Fragment, Antigen-Binding)
★ The Fab region is formed by the arms of the antibody, which adopt a V-shaped
structure designed for binding specific antigens.

★ Structurally, it includes:
 One constant domain and one variable domain from both the heavy and light
chains.
 This combination creates the antigen-binding sites that recognize and attach to
specific molecules.
★ The Fab region is essential for antigen recognition and specificity, as it ensures
antibodies can bind to their corresponding antigens with high precision.
2. Fv Region (Fragment Variable)
★ The Fv region is the smallest functional unit responsible for antigen binding.
★ It is located at the amino-terminal end of the antibody and includes the variable
domains of both the heavy (VH) and light (VL) chains.
★ The antigen-binding site, also called the paratope, is formed by this region. It interacts
directly with the epitope of the antigen.
★ Within the Fv region, the complementarity-determining regions (CDRs), which are
flexible loops, play a critical role in determining antigen-binding specificity.
 CDRs are highly variable and allow the antibody to recognize a wide range of
antigens.
3. Fc Region (Fragment, Crystallizable)
★ The Fc region forms the tail of the antibody and consists of the constant heavy chain
domains (CH2 and CH3 in IgG, IgA, and IgE; CH2, CH3, and CH4 in IgM and IgE).
★ The Fc region is crucial for activating the immune system by interacting with:
 Fc receptors on immune cells such as macrophages, natural killer (NK) cells, and
neutrophils.
  Proteins of the complement system, initiating complement activation and
promoting immune responses such as opsonization and cell lysis.
★ Mediates effector functions like:
 Phagocytosis: Enhancing the uptake of antibody-coated antigens by immune
cells.
 Cytotoxicity: Directing cytotoxic immune cells to destroy pathogens or infected
cells.
 Immune clearance: Facilitating the removal of immune complexes from the
bloodstream.
4. Complementary Roles
★ The Fab and Fv regions specialize in antigen recognition and binding, ensuring
antibodies can identify specific threats.

★ The Fc region bridges antigen recognition with immune system activation, enabling a
coordinated response to neutralize or eliminate the antigen.
★ Together, these regions allow antibodies to:
 Bind specifically to antigens using the Fab region and paratope.
 Activate immune mechanisms via the Fc region, triggering responses such as
inflammation, cell recruitment, and destruction of pathogens.

Fc and Fab portions of Antibody

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 Classes of antibodies
B lymphocytes produce the glycoproteins known as antibodies, or immunoglobulins (Ig),
which are essential to the immune response because they can recognise and attach to particular
antigens. As you said, there are five different kinds of antibodies: IgG, IgM, IgA, IgD, and IgE.
The effector functions and heavy chain structure of these classes vary. Their Fc regions (the
constant region) and heavy chains differ, which results in different characteristics and functional
roles even though they have a fundamental four-chain structure (two heavy chains and two light
chains).

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Immunoglobulin G – IgG
The most prevalent immunoglobin, IgG, makes up almost 80% of all serum antibodies.
IgG levels in the blood are roughly 10 mg/ml.
Structure
The basic structure of IgG is composed of a Y-shaped protein where the Fab arms are
linked to the Fc arms by an extended region of polypeptide chain called the hinge.The region is
exposed and sensitive to attack by proteases that cleave the molecule into distinct functional
units arranged in a four-chain structure.An IgG molecule consists of two identical γ heavy
chains, usually of the size 50kDa. The light chains in IgG exist in two forms; κ and λ, where the
κ form is more prevalent than λ, in the case of humans.

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Characteristic features of IgG :

The monomeric IgG antibodies found in serum can pass from the mother to the foetus
across the placenta. IgG antibodies attach to two distinct epitopes on two distinct antigens
through two paratopes. Based on the γ heavy chain subclasses, IgG is divided into four
subclasses. IgG antibodies are primarily involved in secondary immunological responses, which
are produced as a consequence of response maturation and class switching.

Subclasses of IgG:
Three subclasses of IgG antibodies have been identified: IgG1, IgG2, IgG3, and IgG4.

IgG1
It is the most prevalent of them, and they are called according to their serum quantity.
The most prevalent subclass of IgG antibodies with γ1 heavy chains is IgG1. Although soluble
protein antigen and membrane proteins are the main factors that produce IgG1, smaller amounts
of the other subclasses are frequently present as well. Since IgG1 is the most prevalent subtype, a
lack of it can result in a lower level of total IgG.

IgG2.
Composed of γ2 heavy chains, IgG2 is the second most prevalent kind of IgG in human
serum. Nearly all of the reaction to bacterial capsular polysaccharide antigens is mediated by
IgG2. The only subtype of IgG antibodies that is unable to pass through the placenta during
pregnancy is IgG2. IgG2 deficiency may lead to a weakened immune response to harmful
microbes.

IgG3
The third most prevalent kind of IgG seen in human serum is IgG3, which has γ3 heavy
chains. These are especially good at causing effector functions to be triggered. It is a short-lived,
strong pro-inflammatory antibody. With a strong affinity for FcR on phagocytic cells, IgG3 is
also the most efficient complement activator and facilitates opsonisation.
IgG4
With γ4 subclasses of heavy chains, IgG4 is the least prevalent IgG antibody in human
serum. IgG4 is produced in response to repeated or prolonged exposure to an antigen in a non-
infectious environment and is triggered by allergens. IgG4 has the ability to go from the mother
to the foetus across the placenta. Despite the rarity of IgG4 deficits, elevated serum IgG4 levels
have been linked to several issues in several organs.

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Functions of IgG (Immunoglobulin G)
It is the most abundant and versatile antibody in the blood and extracellular fluid. It
performs a wide range of functions in the immune response due to its structural properties.
1. Neutralization of Pathogens and Toxins
★ IgG can bind to pathogens (such as viruses and bacteria), preventing them from infecting
host cells. This is especially important in blocking viruses from attaching to cell
receptors.
★ IgG can also neutralize bacterial toxins by binding to them, thus preventing their harmful
effects on host tissues.

Neutralization of Pathogens and Toxins

Image source: Abbas et al., Cellular and Molecular Immunology, 7e
2. Opsonization
★ Enhancement of Phagocytosis: IgG antibodies can bind to the surface of pathogens and
mark them for destruction by immune cells such as macrophages and neutrophils. This
process is known as opsonization. The Fc region of IgG interacts with Fc receptors on
phagocytes, enhancing their ability to engulf and digest pathogens.

Opsonisation
Image source: Abbas et al., Cellular and Molecular Immunology, 7e

3. Complement Activation
★ IgG can initiate the classical complement cascade by binding to antigens on the surface of
pathogens. When IgG binds to an antigen, it undergoes a conformational change that
allows it to interact with the first component of the complement system (C1q), leading to
the activation of the complement cascade.
★ Complement activation results in the formation of the membrane attack complex (MAC),
which can directly lyse pathogen cells, and the production of opsonins like C3b, further
enhancing phagocytosis.
4. Antibody-Dependent Cellular Cytotoxicity (ADCC)
★ IgG antibodies can bind to infected or tumor cells and recruit immune cells like natural
killer (NK) cells.
★ NK cells have Fc receptors that bind the Fc region of IgG, triggering the release of
cytotoxic molecules that kill the target cell. This is particularly important in fighting viral
infections and cancerous cells.
 Antibody-Dependent Cellular Cytotoxicity (ADCC)
Image source: Abbas et al., Cellular and Molecular Immunology, 7e

5. Placental Transfer
★ IgG is the only antibody class that can cross the placenta, providing passive immunity to
the developing fetus. This transfer occurs through a receptor on placental cells, which
allows IgG to be transported across the placenta into the fetal bloodstream. This provides
the newborn with immediate protection against infections during the early months of life
until it’s immune system matures.
6. Regulation of Immune Responses
★ IgG antibodies also play a role in regulating the immune response. They can interact with
immune cells to fine-tune the production of other immune mediators. For instance,
certain IgG subclasses can influence T cell responses and modulate the activity of other
immune cells.
7. Memory Response in Adaptive Immunity
★ IgG is a major component of the immune memory generated after an infection or
vaccination. After initial exposure to a pathogen, B cells produce IgG antibodies, and
subsequent exposures result in a faster, more robust IgG-mediated immune response. IgG
is typically the predominant antibody produced during secondary immune responses,
which is key to long-lasting immunity.
 Immunoglobulin M (IgM)
IgM is the third most abundant immunoglobulin in the serum, with a concentration of
approximately 1.5 mg/ml. It is the largest antibody and is the first to be produced during the
body's initial response to an antigen.

Structure of IgM
IgM is secreted primarily in a pentameric form, consisting of five subunits, each made up
of two µ heavy chains and two light chains. In some cases, a J chain may be present in the
hexameric form of the molecule, though it is not always included. The J chain is typically added
just before secretion, as it facilitates the polymerization of the individual monomers. Each
monomer contains two antigen-binding sites, giving the entire pentamer 10 binding sites.
However, not all of these binding sites can be occupied simultaneously due to spatial constraints.
The pentameric IgM has a molecular weight of 900 kDa.

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Properties of IgM
IgM is the largest antibody in the human body and the only antibody that exists in a
pentameric form. It is the first antibody produced in response to an initial exposure to an antigen,
marking the early stages of the immune response.
✿ IgM is the first immunoglobulin produced by the fetus, typically beginning around the
20th week of gestation. This early production plays a crucial role in the immune defense
of the developing fetus.
✿ IgM is composed of five monomeric subunits, each containing two heavy chains (µ) and
two light chains. These monomers are connected by disulfide bonds, forming a pentamer
with a total of 10 antigen-binding sites. This large structure allows IgM to bind multiple
antigens simultaneously, enhancing its ability to neutralize pathogens.
✿ In its monomeric form, IgM functions as the primary antibody receptor on the surface of
B lymphocytes. This form plays a key role in initiating the immune response when it
binds to specific antigens.
✿ Compared to other antibodies like IgG, IgM has a relatively short lifespan. It is typically
produced early in the immune response but is replaced by other antibody classes like IgG
over time, which provide longer-lasting immunity.
✿ The large size of IgM molecules restricts their ability to diffuse effectively across tissues.
As a result, IgM is found in relatively low concentrations in intracellular fluids, such as
the cytoplasm. It is more abundant in the bloodstream and other extracellular fluids.
Functions of IgM
IgM plays several crucial roles in the immune response, especially during the early stages
of an infection. Its structure and properties make it highly effective against various pathogens,
particularly viruses and bacteria.
✿ Effective Against Viruses: IgM is particularly effective in neutralizing viral infections.
Compared to IgG, a smaller amount of IgM is required to neutralize viruses, making it an
essential antibody during the initial immune response. Its large size and pentameric
structure allow it to bind tightly to viral particles, preventing them from infecting host
cells.
✿ Superior Agglutination Properties: IgM has much stronger agglutination properties
than other antibodies like IgG. In fact, it takes between 100 to 1000 times more IgG
 molecules than IgM to achieve the same level of agglutination. This is due to the
pentameric structure of IgM, which allows it to cross-link multiple antigens
simultaneously, leading to a more effective clumping (agglutination) of pathogens or
infected cells. Agglutination helps in the clearance of these pathogens by immune cells.
✿ Complement Activation via the Classical Pathway: IgM is a potent activator of the
classical pathway of the complement system, an essential part of the immune defense.
The presence of two Fc regions in close proximity within the IgM pentamer enables it to
interact effectively with the first component of the complement system (C1q). When IgM
binds to antigens on the surface of pathogens, it triggers a cascade of events in the
complement system, leading to the formation of the membrane attack complex (MAC)
and the destruction of the pathogen. This enhances the immune system's ability to clear
infections.

Immunoglobulin A (IgA)
IgA, also known as sIgA (secretory IgA), is a crucial immunoglobulin primarily found in
mucosal areas of the body, where it functions as a protective antibody in various secretions.
Though present in relatively low concentrations in the blood, IgA is found in much higher
concentrations in bodily fluids like tears, saliva, sweat, and mucus. Its main role is to safeguard
mucosal surfaces from pathogens and foreign particles.

Structure of IgA

IgA has a molecular weight of around 160 kDa and is usually present as a monomer.
However, it can also exist in dimeric or trimeric forms, particularly in mucosal areas where it is
secreted to provide enhanced protection. The structure of IgA consists of four chains: two heavy
chains and two light chains.
 Structure of dimeric IgA
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✿ Heavy Chain Structure: The heavy chain of IgA is divided into three constant domains
(CH1, CH2, and CH3) and a variable region (VH) that is responsible for antigen
recognition.
✿ Hinge Region: The hinge region, located between the CH1 and CH2 domains, is a
flexible section that helps the antibody adapt its shape to better bind to antigens. This
region is held together by disulfide linkages, providing structural stability.
✿ Secretory Component: One of the defining features of sIgA is its secretory component,
which is an additional polypeptide chain of 75 kDa. This component helps in the
transport of IgA across epithelial cells and protects the antibody from degradation by
enzymes in bodily fluids, such as proteases.
✿ J Chain: IgA also contains a J-chain, which links the individual monomers in the dimeric
or trimeric form via disulfide bridges. The J-chain plays an important role in
polymerization and facilitates the secretion of IgA into mucosal surfaces, where it can
exert its protective function.
 ✿ IgA is divided into two subclasses: IgA1 and IgA2.
 IgA1 exists primarily as a monomer and makes up about 80% of the IgA
found in serum. It is produced in the bone marrow and released onto mucosal
surfaces.
 IgA2, on the other hand, is typically found in polymeric forms, including
dimeric structures, and is more commonly present in secretions produced
locally.
 A key difference between IgA1 and IgA2 is the structure of their hinge
regions. In IgA1, the hinge region is notably longer and more extended
compared to IgA2.

Detailed Structure of IgA

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Function of IgA
★ Mucosal Immunity: IgA is predominantly found in mucosal surfaces such as the
respiratory, gastrointestinal, and urogenital tracts, as well as in secretions like breast milk,
tears, and saliva. It serves as a first line of defense by preventing the attachment of
pathogens (like bacteria and viruses) to epithelial cells, thus blocking infections at these
entry points.
 ★ Barrier Protection: In its secretory form (sIgA), it forms a protective barrier against
harmful microorganisms and particulate antigens, neutralizing them and preventing their
penetration through mucosal membranes.
★ Protection from Enzymatic Degradation: The secretory component and the J-chain
confer protection to IgA against proteolytic enzymes that are abundant in mucosal
environments, ensuring that IgA maintains its functional integrity while performing its
immune functions.

Functions of IgA
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Immunoglobulin D (IgD)
IgD is a monomeric antibody primarily found on the surface of immature B
lymphocytes, where it functions as a receptor for antigens. It is produced in small amounts and
secreted into the bloodstream, but its concentration in serum is much lower than other
immunoglobulins like IgG and IgM.
Structure of IgD
IgD exhibits a significant degree of structural diversity across vertebrate species,
adapting to fulfill its role alongside IgM in the immune system. Its flexibility is crucial for
complementing the functions of other antibodies, particularly IgM.
✿ Basic Structure: IgD is a glycoprotein made up of two identical δ (delta) heavy chains
and two identical light chains. These chains are linked by disulfide bonds, forming the
classic Y-shaped structure seen in immunoglobulins.
✿ Membrane Anchoring: The IgD that is found on the surface of B lymphocytes has
additional amino acid sequences at the C-terminal region, which anchor it to the cell
membrane. This membrane-bound form is essential for its role as an antigen receptor on
B cells.
✿ Disulfide Linkages and Domains: The light and heavy chains of IgD are connected by
interchain disulfide bonds. Additionally, there are intrachain disulfide bonds that
divide the chains into different structural domains. These domains contribute to the
stability and flexibility of the IgD molecule.
✿ Extended Hinge Region: One distinctive feature of IgD is its extended hinge region,
which provides increased flexibility to the antibody. This flexibility allows IgD to better
interact with antigens. However, the longer hinge region also makes IgD more
susceptible to proteolytic cleavage, which may limit its durability under certain
conditions.

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Properties of IgD
IgD is a relatively rare antibody in terms of both its serum concentration and its specific
functions within the immune system. While its precise role remains somewhat unclear, it has
distinct properties that contribute to its importance in B cell activation and immune responses.
✿ IgD is found in very low concentrations in the bloodstream, making up only about
0.25% of the total serum immunoglobulins. Despite its low serum levels, it plays a key
role in certain aspects of immune function, particularly in its membrane-bound form.
✿ IgD has a relative molecular mass of 185 kDa, and its half-life in circulation is
approximately 2.8 days. This relatively short half-life reflects its transient presence in the
bloodstream, as it is primarily found as a receptor on B cells rather than in free
circulation.
✿ Around 1% of the proteins found in the plasma membranes of B lymphocytes are IgD.
On these B cells, IgD functions as a key receptor for antigens. It is typically co-
expressed with IgM on the surface of B lymphocytes, where both antibodies contribute
to the initiation of immune responses. This co-expression suggests that IgD works in
concert with IgM to facilitate the activation and differentiation of B cells, aiding in their
response to pathogens.
Functions of IgD
IgD, though less studied compared to other immunoglobulins, plays a pivotal role in the
immune system, particularly in B cell function and regulation. While its exact functions are
still being clarified, several key roles have been identified that are crucial for immune responses,
especially in the early stages of immune activation.
1. B Cell Activation and Antigen Receptor: One of the primary functions of IgD is its role
as a B cell receptor (BCR). It is expressed on the surface of immature and mature B
lymphocytes, where it functions in concert with IgM to detect and bind antigens. This
binding triggers the activation of B cells, leading to their differentiation and the
production of antibodies. IgD, as a receptor, helps initiate the adaptive immune response
by recognizing foreign antigens, especially during the early phases of immune
surveillance.
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2. Co-expression with IgM: On the surface of B cells, IgD is often co-expressed with
IgM, both of which serve as receptors for antigens. This co-expression ensures that B
cells have multiple ways of recognizing pathogens. IgM is more efficient at initiating
immune responses, but IgD plays a complementary role in fine-tuning the response.
Together, these antibodies collaborate to regulate the activation and differentiation of B
cells.
3. Regulation of B Cell Tolerance: IgD has been suggested to play a role in immune
tolerance, helping to prevent B cells from attacking the body's own tissues. This function
is essential for maintaining immune homeostasis and preventing autoimmune reactions. It
is thought that IgD may influence the selection and survival of B cells in the bone
marrow, ensuring that only B cells capable of recognizing foreign pathogens, rather than
self-antigens, are allowed to mature and enter circulation.
4. Activation of Signaling Pathways: When IgD binds to an antigen, it activates
intracellular signaling pathways in B cells, leading to changes in gene expression that
promote B cell activation, survival, and proliferation. This signaling is crucial for
initiating the adaptive immune response and ensuring that the immune system can mount
an appropriate response to infections.
5. Mucosal Immunity: While IgD is primarily found in the blood and on B cell surfaces, it
may also have a role in mucosal immunity, where it can participate in the local immune
 responses within mucosal tissues, such as those in the respiratory and gastrointestinal
tracts. Some evidence suggests that IgD may help in the defense against pathogens by
interacting with mucosal epithelia, though its role in this context is less well understood.
6. Facilitation of B Cell Memory Formation: IgD is thought to play a role in the
development of B cell memory. After the initial antigen encounter, IgD may help in the
differentiation of B cells into long-lived memory cells. These memory B cells are crucial
for providing rapid and robust immune responses upon subsequent exposure to the same
pathogen.

Immunoglobulin E (IgE)
IgE (Immunoglobulin E) is a class of antibody primarily involved in the body’s immune
response to allergens and parasitic infections. It is present in very low concentrations in the
blood, but its functions are critical, especially in allergic reactions and defense against parasitic
organisms such as worms. IgE is best known for its role in hypersensitivity reactions, including
asthma, hay fever, and anaphylaxis. When an individual is exposed to an allergen, IgE binds to
the surface of mast cells and basophils through high-affinity receptors (FcεRI), leading to the
release of histamines and other inflammatory mediators upon subsequent exposures, to the same
allergen.
Structure of IgE
IgE, like other antibodies, has a basic Y-shaped structure consisting of two heavy
chains and two light chains. However, there are specific features that differentiate IgE from
other immunoglobulins, such as IgG, IgA, and IgM.
✿ Heavy Chains: IgE is composed of two heavy chains labeled ε (epsilon). These heavy
chains are longer than those found in other immunoglobulins, and they contain four
constant domains (CH1, CH2, CH3, and CH4). The CH3 and CH4 domains of the
heavy chain play key roles in binding to the high-affinity FcεRI receptors on mast cells
and basophils.
✿ Light Chains: Like other antibodies, IgE has two light chains, which can be of either
kappa (κ) or lambda (λ) type, and they are linked to the heavy chains by disulfide
bonds.
 ✿ Hinge Region: The hinge region in IgE provides flexibility to the molecule, allowing it
to interact with receptors and bind to antigens. This region is somewhat different from
that of IgG and IgA antibodies, contributing to the specific functional interactions of IgE.
✿ Fc Region: The Fc region (the tail of the Y-shaped antibody) is a crucial part of IgE's
structure. It interacts specifically with the FcεRI receptor found on mast cells and
basophils. This interaction is what enables IgE to trigger the release of histamine and
other inflammatory mediators when an allergen binds to the IgE on these cells.
✿ Antigen-Binding Site: The Fab region (the arms of the Y-shaped antibody) contains the
antigen-binding sites, where IgE can specifically bind to allergens or parasitic antigens.
The variable regions of the heavy and light chains are responsible for recognizing and
binding to these antigens, initiating the immune response.

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Properties of IgE
IgE is an essential immunoglobulin in the immune system, primarily involved in allergic
reactions and defending against parasitic infections. Although it is present in very low
concentrations in the bloodstream, its effects are potent, especially in triggering inflammation
and immune responses to allergens.
1. IgE is found in extremely low concentrations in the blood, accounting for only about
0.05% of the total immunoglobulins in the serum. Despite this, IgE has significant
 biological activity due to its role in hypersensitivity reactions and defense against
parasitic organisms.
2. One of the most distinctive features of IgE is its **high affinity for binding to the FcεRI
receptor on mast cells and basophils. When IgE binds to these cells, it "primes" them to
react more rapidly upon subsequent exposure to the same antigen (allergen). This
interaction is crucial for the immediate hypersensitivity response seen in allergic
reactions. Upon re-exposure to the allergen, IgE bound to mast cells or basophils triggers
these cells to release histamine, cytokines, and other inflammatory mediators that cause
the symptoms of an allergic reaction.
3. IgE is the central antibody in allergic responses. When an individual encounters an
allergen for the first time, IgE is produced and binds to the surfaces of mast cells and
basophils. In subsequent exposures, the allergen binds to the IgE, triggering
degranulation of these cells and the release of chemicals like histamine, which lead to
the symptoms of allergic reactions, such as swelling, itching, and difficulty breathing (in
severe cases, anaphylaxis).
4. IgE also plays a key role in defending against parasitic infections, particularly those
caused by helminths (worms). During a parasitic infection, IgE is produced and binds to
the surface of the parasite. This binding activates immune cells, including eosinophils,
which release toxic granules that help to destroy the parasite. IgE thus contributes to
immune defense against parasitic invaders by recruiting and activating effector cells that
can directly damage the pathogen.
5. IgE has a relatively short half-life in circulation compared to other immunoglobulins,
typically around 2 to 3 days. However, its binding to mast cells and basophils prolongs
its activity, as it remains on these cells until it is triggered by antigen exposure.
6. IgE is predominantly found on the surface of mast cells and basophils, rather than
circulating freely in the blood. This restricted distribution is a key factor in its role in
localized immune responses, especially in the tissues of the skin, lungs, and
gastrointestinal tract, where allergic reactions commonly occur.
7. IgE’s functions are distinct from those of other immunoglobulin classes like IgG, IgM,
and IgA. It is specifically adapted to promote allergic responses and parasitic
 immunity, unlike other antibodies, which are involved in different immune functions like
bacterial opsonization (IgG), mucosal immunity (IgA), and complement activation (IgM).
8. Due to its central role in allergies, IgE is a target in immunotherapy for allergic
conditions. Anti-IgE therapies (e.g., omalizumab) are used to block IgE binding to
mast cells and basophils, thus preventing the release of histamines and other
inflammatory mediators. This approach helps in treating conditions like asthma and
chronic urticaria (hives).

Functions of IgE
Immunoglobulin E (IgE) is a specialized antibody that plays a crucial role in the body’s
response to allergens and parasitic infections. Though it is present in very low concentrations
in the bloodstream, its functional importance is significant, especially in mediating allergic
reactions and in immune responses against parasites.
1. Mediating Allergic Reactions
IgE is central to the immune response in allergic diseases. When a person with an
allergy encounters an allergen for the first time, the immune system produces IgE antibodies
specific to that allergen. This IgE binds to the surface of mast cells and basophils, which are
immune cells involved in inflammation.
 Sensitization: The first exposure to an allergen leads to IgE production specific to that
allergen. This IgE binds to FcεRI receptors on the surfaces of mast cells and basophils,
"sensitizing" them.
 Subsequent Exposure and Degranulation: Upon later exposure to the same allergen,
the allergen binds to the IgE on the mast cells or basophils. This triggers cell
degranulation, releasing histamines, cytokines, and other pro-inflammatory mediators.
 Allergic Symptoms: The release of these chemicals leads to the classic symptoms of
allergic reactions such as itching, swelling, redness, rashes, and respiratory issues (like
sneezing, wheezing, or anaphylaxis). For example, in hay fever, asthma, and hives, IgE-
mediated responses contribute significantly to the severity of the symptoms.
 Anaphylaxis: In severe cases, IgE activation can lead to anaphylaxis, a life-threatening
allergic reaction that requires immediate medical intervention. The widespread release of
inflammatory mediators can cause severe vasodilation, bronchoconstriction, and shock.
 IgE Mediated allergic response
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2. Defense Against Parasitic Infections

IgE also plays an important role in defending the body against parasitic infections,
particularly those caused by helminths (worms), such as hookworms, roundworms, and
tapeworms.
 Immune Activation: When IgE binds to antigens present on the surface of parasites,
it triggers an immune response that activates various immune cells, particularly
eosinophils and mast cells.
 Eosinophil Activation: Eosinophils are a type of white blood cell that contains toxic
granules. IgE binds to the parasite, leading to the recruitment and activation of
eosinophils. These cells release toxic proteins, such as major basic protein, which
are capable of damaging and killing the parasitic invader.
 Mast Cells and Basophils in Parasite Defense: Mast cells and basophils, upon
activation by IgE, release cytokines and other inflammatory mediators that help
 coordinate the immune response against the parasite. The goal is to expel the parasite
from the body and limit its damage.
 Th2 Response: IgE is particularly involved in promoting a Th2-type immune
response, which is associated with increased eosinophil activity and the production of
IgE, facilitating the immune defense against larger parasites.
3. Immunoregulation and Immune Memory
While IgE is primarily known for its involvement in allergy and parasitic defense, it also
plays a role in immune regulation.
 Th2 Cytokine Production: IgE-mediated responses promote the production of Th2
cytokines (like IL-4, IL-5, and IL-13), which further enhance the allergic response and
also regulate eosinophil recruitment during parasitic infections.
 Memory Response: Like other antibodies, IgE can participate in the formation of
immune memory, ensuring a quicker and stronger immune response when the body is
re-exposed to the same allergen or pathogen. This is why individuals who have been
sensitized to an allergen can experience increasingly severe reactions upon re-exposure.
4. Protective Role in Mucosal Immunity
Although IgE is primarily associated with systemic allergic reactions, it may also play a
role in mucosal immunity—protecting the body's mucosal surfaces (such as those in the
respiratory and gastrointestinal tracts) from pathogens and parasites.
 Local Responses: In some cases, IgE may help orchestrate localized immune responses
in mucosal tissues, including the respiratory tract (in conditions like allergic rhinitis or
asthma) or the gastrointestinal tract (in defense against parasitic worms).
 Barrier Protection: By promoting the activation of mast cells and eosinophils at
mucosal surfaces, IgE helps enhance the first line of defense against allergens, pathogens,
and parasites, facilitating rapid immune responses at sites of entry.
5. Modulation of Immune System Homeostasis
IgE also plays a less-well-understood but important role in modulating immune system
balance. For instance, while it contributes to allergic disease, IgE may also help maintain
balance within the immune system by regulating Th2-type immune responses, which are
involved in combating helminth infections. In certain conditions, this balance is disrupted,
contributing to allergic diseases and autoimmunity.
6. IgE in Immunotherapy
Due to its central role in allergic diseases, IgE has become a therapeutic target for
immunotherapy in treating conditions like asthma, chronic urticaria (hives), and other allergic
disorders. For example:
 Anti-IgE Therapy: Drugs like omalizumab (Xolair) are monoclonal antibodies that bind
to IgE and prevent it from attaching to mast cells and basophils. This prevents IgE-
mediated degranulation and the release of histamine and other inflammatory mediators,
thereby reducing allergic reactions and improving the symptoms of allergic diseases.
 hybridoma technology and Monoclonal
antibodies
Introduction:
Hybridoma technology is used to produce monoclonal antibodies. The term hybridoma
refers to the hybrid cells formed by fusing two different types of cells: a lymphocyte and a tumor
cell. One of the cells involved is a B-lymphocyte, such as a spleen cell from a mouse that has
been immunized with sheep red blood cells. The other is a single myeloma cell (a type of bone
marrow tumor cell) that can grow indefinitely in culture. The fusion of these two cells combines
their abilities: the lymphocyte's capacity to produce large amounts of pure antibodies and the
tumor cell's ability to proliferate endlessly. This results in hybridoma cells that can continuously
generate the desired monoclonal antibodies.
Monoclonal antibodies (mAbs) are a type of antibody that is specifically produced to
target a single antigen. Normally, antibodies are glycoproteins produced in the bloodstream to
defend against foreign antigens and provide immunity. These naturally occurring antibodies are
polyclonal, meaning they are a mix of various antibodies that target different parts of an antigen,
and thus lack specificity.
In contrast, when a specific lymphocyte is isolated and cultured in the lab, and is capable
of producing only one type of antibody that targets a specific antigen, this is called a monoclonal
antibody. Monoclonal antibodies are derived from a single cell clone, and they are designed to
recognize and bind to a particular antigen. These antibodies can be produced against a wide
range of substances, including proteins, glycoproteins, glycolipids, nucleic acids, and other
chemically defined groups that are linked to protein carriers.

Steps of Hybridoma Technology:

Step I: Immunization of rabbit or rat and extraction of B-lymphocytes: To isolate the B-
lymphocytes that produce specific antibodies, a rabbit or lab rat is first immunized through
multiple injections of a specific antigen, such as sheep red blood cells (RBCs). After
immunization, B-cells are collected from the spleen of the animal.
Step II: Fusion of myeloma cells with B-lymphocytes: The extracted B-lymphocytes are then
mixed with myeloma cells, which are derived from bone marrow. The goal is to create
hybridoma cells through the fusion of the B-cells and myeloma cells. This fusion can be
 achieved using various methods, such as polyethylene glycol (PEG), electroporation, or
phage-mediated fusion.
Step III: Selection of Hybridoma Cells: The next step involves selecting the hybridoma cells.
B-lymphocytes contain the HPRT1 gene, which codes for the enzyme hypoxanthine-
guanine phosphoribosyltransferase (HGPRT). This enzyme is essential for synthesizing
nucleotides from hypoxanthine, which is present in the culture medium. As a result, B-cells
can grow in a medium containing Hypoxanthine, Aminopterin, and Thymine (HAT
medium).
However, myeloma cells lack the HPRT1 gene, so they do not produce the
HGPRT enzyme and cannot survive in HAT medium. Myeloma cells that fuse with other
myeloma cells, or those that don't fuse at all, will die because they can't use hypoxanthine.
Similarly, B-cells that fuse with other B-cells or do not fuse at all will also die eventually, as they
lack the ability to divide indefinitely. Therefore, only the hybridoma cells formed by the fusion
of B-cells and myeloma cells can survive and proliferate in HAT medium. The hybridoma cells
are then screened and selected as the desired cells for monoclonal antibody production.
Step IV: Culturing Hybridoma Cells: The selected hybridoma cells are then cultured in an
appropriate culture medium, which is often supplemented with substances like insulin,
transferrin, ethanol, amines, and other hormones. Commonly used culture media for
hybridoma cells in monoclonal antibody production include:
 DMEM (Dulbecco's Modified Eagle Medium)
 IMDM (Iscove's Modified Dulbecco's Medium)
 Ham's F12
 RPMI 1640 medium (Roswell Park Memorial Institute 1640 Medium)
Step V: Inoculation of Hybridoma Cells into a Suitable Host: The hybridoma cells are then
injected into a lab animal, where they begin to produce monoclonal antibodies. These hybridoma
cells can also be frozen and stored for future use.
Step VI: Extraction and Purification of Monoclonal Antibodies: Monoclonal antibodies are
extracted from the host animal and purified using one of the following methods:
 Ion exchange chromatography
 Antigen affinity chromatography
 Radial immunoassay
  Immune precipitation

Steps in Hybridoma technology

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production.jpg

Types of Monoclonal Antibodies based on their origin:

Monoclonal antibodies are artificially created to mimic human antibodies in the immune
system and are produced in four main ways, each named based on their origin.
A. Murine Monoclonal Antibodies: Murine monoclonal antibodies were the first to be
produced on a laboratory scale using hybridoma technology in 1975. They are called
"murine" because they are derived from rodent hosts (mice and rats) in the Muridae
family. These antibodies were initially tested in preclinical trials for treating
Cryptococcus neoformans. Murine mAbs played a significant role in advancing modern
antibody production techniques and their potential uses in both therapeutics and
analytical applications. The names of treatments using murine mAbs typically end with "-
omab."
B. Chimeric Monoclonal Antibodies: Chimeric monoclonal antibodies are hybrids made by
combining the variable regions from mice with the constant regions from humans. These
chimeric antibodies were developed to reduce immunogenicity and increase serum half-
life, making them more suitable for therapeutic use. Chimeric mAbs maintain the original
 antibody’s antigen specificity and affinity. Treatments using chimeric mAbs are named
with the suffix "-ximab."
C. Humanized Monoclonal Antibodies: Humanized monoclonal antibodies are a further
development of chimeric monoclonal antibodies. In humanized mAbs, all parts of the mouse
antibody are replaced with human components, except for the complementarity-determining
regions (CDRs), which are the amino acids responsible for directly interacting with the
antigen. This means that humanized mAbs contain small segments of mouse protein attached
to the human antibody. Treatments using humanized mAbs typically end with the suffix "-
zumab."
D. Human Monoclonal Antibodies: Human monoclonal antibodies are completely human
proteins that have been modified using molecular biology techniques to alter their amino acid
sequences. This modification changes their specificity, affinity, or biological function,
incorporating sequences that are not normally found in the human genetic code. Treatments
using fully human mAbs end with the suffix "-umab."

Types of Monoclonal Antibodies based on their origin

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Types of Monoclonal Antibodies Based on Function: Monoclonal antibodies can also be

categorized according to the roles they play, such as those used in cancer treatment:
1. Naked Monoclonal Antibodies: Naked monoclonal antibodies (mAbs) are not linked to any
drugs or radioactive agents. These are among the most common mAbs used in cancer
therapy. Most naked mAbs bind to antigens on cancer cells, while others may attach to
 antigens on non-cancerous cells or free-floating proteins. These naked mAbs work in various
ways, such as:
a) Enhancing the immune response against cancer cells by binding to the cancer cells
and acting as signals for the immune system to target and destroy them. For example,
Alemtuzumab (Campath®), used for treating Chronic Lymphocytic Leukemia (CLL),
binds to the CD52 antigen on lymphocytes, including leukemic cells, thereby attracting
immune cells to eliminate them.
b) Targeting immune system checkpoints to boost the immune response and improve the

body’s ability to fight cancer.

c) Blocking antigens on cancer cells that promote the growth and spread of cancer. An
example is Trastuzumab (Herceptin), which targets the HER2 protein on breast and
stomach cancer cells. By blocking this protein, it prevents the growth and spread of
cancer cells.
2. Conjugated (Labeled, Tagged, or Loaded) Monoclonal Antibodies: Conjugated
monoclonal antibodies are combined with chemotherapy drugs or radioactive agents to
enhance targeted treatment. These mAbs act as "homing devices," delivering chemotherapy
directly to the cancer cells. Conjugated mAbs circulate throughout the body until they attach
to the target antigen, where they deliver the attached drug or agent to initiate immune
responses for elimination of the cancer cells. The advantage of conjugated mAbs is that they
help minimize the damage to healthy cells in other parts of the body.
a. Radiolabeled Monoclonal Antibodies: Radiolabeled mAbs are linked to a small radioactive
particle. These antibodies deliver both the drug and the radioactive agent directly to the target
cells, causing damage not only to the cancer cells but also to nearby cells. For example,
Ibritumomab tiuxetan (Zevalin) targets the CD20 antigen on B-lymphocytes and delivers
radioactivity directly to cancer cells. Ibritumomab tiuxetan is a combination of the
monoclonal antibody rituximab and a radioactive agent, Yttrium-90. This treatment is
referred to as radioimmunotherapy (RIT).
b. Chemolabeled Monoclonal Antibodies: These mAbs are attached to powerful chemotherapy
drugs or other therapeutic agents.
3. Bispecific Monoclonal Antibodies: Bispecific monoclonal antibodies are drugs that combine
two different monoclonal antibodies linked together. For example, Blinatumomab
 (Blincyto), used for leukemia, has one part that targets the CD19 protein on leukemia and
lymphoma cells and another part that binds to the CD3 protein on immune T-cells.

Functions of MAbs:
Monoclonal antibodies (mAbs) have a wide range of functions depending on how they
are designed and what they target. These functions are leveraged in diagnostics, therapeutics, and
research. Some of the important functions of monoclonal antibodies:
1. Targeted Immune Response: One of the primary uses of mAbs is to enhance the immune
system’s ability to recognize and destroy cancer cells, pathogens, or other harmful entities.
★ Immune System Activation: mAbs can bind to cancer cells or infected cells and act as a

signal to the immune system, marking them for destruction. This process is called
antibody-dependent cellular cytotoxicity (ADCC), where immune cells such as natural
killer (NK) cells or macrophages recognize and destroy the antibody-coated cells.
★ Complement Activation: Some mAbs can activate the complement system, a part of the

immune system that enhances the ability of antibodies and phagocytic cells to clear
pathogens or damaged cells. This process is known as complement-dependent
cytotoxicity (CDC).
★ Checkpoint Inhibition: Certain mAbs target immune checkpoint proteins (like PD-1 or

CTLA-4) that normally inhibit immune responses. By blocking these checkpoints, mAbs
can help “unmask” cancer cells, allowing immune cells to attack them more effectively.

2. Direct Tumor Cell Killing: Some monoclonal antibodies are designed to bind directly to
tumor cell markers and induce cell death.
★ Blocking Growth Signals: mAbs can block the binding of growth factors to their

receptors on cancer cells. This prevents the cancer cells from receiving the signals that
promote their growth and survival. For example, trastuzumab (Herceptin) targets the
HER2 receptor on breast cancer cells and inhibits their growth.
★ Inducing Apoptosis: Certain mAbs are designed to directly induce apoptosis
(programmed cell death) in cancer cells by binding to specific cell surface proteins. For
instance, rituximab targets the CD20 antigen on B-cells, leading to their destruction and
apoptosis.
3. Delivery of Cytotoxic Agents: Conjugated monoclonal antibodies are used to deliver
cytotoxic agents directly to cancer cells.
★ Radiolabeled mAbs: These antibodies are linked to radioactive isotopes, allowing them

to deliver radiation directly to the tumor, minimizing damage to surrounding healthy

tissue. For example, ibritumomab tiuxetan (Zevalin) is a radiolabeled mAb used in the
treatment of certain lymphomas.
★ Chemolabeled mAbs: These are mAbs attached to chemotherapy drugs, which can

deliver powerful cytotoxic agents directly to the cancer cells. This method, known as
antibody-drug conjugates (ADCs), minimizes the side effects of chemotherapy on healthy
tissues. An example is brentuximab vedotin (Adcetris), which targets CD30 on
lymphoma cells and delivers a toxic chemotherapy agent to the targeted cells.

4. Blocking or Inhibiting Pathogen Activity: mAbs are also used to target infectious agents
like viruses, bacteria, and fungi.
★ Virus Neutralization: mAbs can neutralize viruses by binding to their surface antigens

and preventing them from entering host cells. This is particularly useful in viral infections
such as HIV, Ebola, and COVID-19. For example, casirivimab and imdevimab are
mAbs used to treat COVID-19 by blocking the spike protein of the SARS-CoV-2 virus
from binding to the human ACE2 receptor.
★ Targeting Bacterial Toxins: Certain mAbs can neutralize bacterial toxins, preventing

them from causing damage to the host. For example, Bezlotoxumab is a mAb used to
treat infections caused by Clostridium difficile by neutralizing its toxins.

5. Diagnostic Applications: Monoclonal antibodies are widely used in diagnostics,

particularly in detecting and quantifying specific molecules.
★ Immunoassays: mAbs are commonly used in diagnostic tests like ELISA (enzyme-

linked immunosorbent assay), Western blotting, and immunohistochemistry to detect

the presence of antigens, pathogens, or cancer markers in blood, tissues, or other samples.
 ★ Immunofluorescence and Immunohistochemistry: mAbs can be labeled with
fluorescent or enzyme tags to detect specific proteins in tissue samples. This is especially
useful for diagnosing cancer and infectious diseases.
6. Treatment of Autoimmune Diseases: Monoclonal antibodies are also employed in
treating autoimmune diseases where the immune system attacks the body's own cells.

★ Blocking Autoimmune Targets: mAbs can target and neutralize specific immune
system components that are causing tissue damage. For example, adalimumab
(Humira) is used to treat rheumatoid arthritis by blocking tumor necrosis factor
(TNF), a molecule involved in inflammation.
★ Reducing Inflammation: mAbs that target immune cells or inflammatory cytokines
can reduce chronic inflammation in diseases like inflammatory bowel disease (IBD),
psoriasis, and lupus. For instance, infliximab (Remicade) targets TNF-alpha to help
reduce inflammation in diseases like Crohn’s disease and ulcerative colitis.
7. Gene Therapy and Cellular Therapy: mAbs are also involved in experimental
treatments, such as gene therapy and cellular therapy.
★ Gene Delivery: Some mAbs are engineered to deliver genetic material to specific cells,

potentially allowing for the treatment of genetic diseases.

★ CAR-T Cell Therapy: Chimeric Antigen Receptor T-cell (CAR-T) therapy involves

modifying a patient’s own T-cells to express a receptor that recognizes a specific cancer
antigen. Monoclonal antibodies can be used to select or enhance the T-cells that are most
effective in targeting the cancer cells.
8. Bispecific Antibodies: These are engineered mAbs that can bind to two different antigens
simultaneously. Bispecific antibodies are useful in cancer immunotherapy, where they
can bind one arm to a cancer cell antigen and the other to a T-cell antigen, effectively
bringing immune cells directly to the cancer cells. This enhances the immune response
against tumors.
Applications of MAbs:
Monoclonal antibodies (mAbs) have a wide range of applications in medicine, research,
and diagnostics due to their ability to target specific molecules with high precision. These
applications can be broadly categorized into therapeutic, diagnostic, and research uses. Below is
a detailed note on the various applications of monoclonal antibodies (mAbs):
1. Therapeutic Applications of Monoclonal Antibodies
Monoclonal antibodies are extensively used in the treatment of several diseases,
particularly cancer, autoimmune diseases, infectious diseases, and others. The therapeutic
potential of mAbs has revolutionized medicine by providing highly specific treatment options
with fewer side effects.
A. Cancer Treatment (Oncology)
Monoclonal antibodies have proven to be effective in the treatment of various cancers by
targeting specific antigens on tumor cells.
⬥ Immune Modulation: mAbs can stimulate the immune system to recognize and destroy

cancer cells. For instance, rituximab (Rituxan) targets CD20 on B-cells and is used for
treating non-Hodgkin lymphoma and chronic lymphocytic leukemia (CLL).
⬥ Targeted Therapy: mAbs can bind to specific receptors or proteins on cancer cells,

inhibiting their growth or inducing cell death. For example, trastuzumab (Herceptin)
targets the HER2 receptor, which is overexpressed in some breast and gastric cancers,
and blocks signaling that promotes cancer cell growth.
⬥ Antibody-Drug Conjugates (ADCs): These are mAbs linked to chemotherapy drugs or

other cytotoxic agents, allowing the drug to be delivered directly to the cancer cell.
Brentuximab vedotin (Adcetris) targets the CD30 antigen on lymphoma cells and
delivers a cytotoxic drug.
⬥ Radiolabeled mAbs: mAbs can be conjugated with radioactive isotopes to deliver

targeted radiation directly to the tumor, minimizing damage to surrounding healthy

tissue. Ibritumomab tiuxetan (Zevalin) is an example of a radiolabeled mAb used in
lymphoma treatment.
⬥ Checkpoint Inhibitors: Some mAbs, such as nivolumab (Opdivo) and pembrolizumab

(Keytruda), target immune checkpoint proteins like PD-1 or PD-L1, which are used by
tumors to escape immune surveillance. These mAbs enhance immune responses against
cancer.
B. Autoimmune Diseases
Monoclonal antibodies are also used to treat autoimmune diseases where the immune
system attacks the body’s own tissues.
 ⬥ TNF Inhibitors: mAbs like infliximab (Remicade), adalimumab (Humira), and

etanercept (Enbrel) block tumor necrosis factor-alpha (TNF-α), a cytokine involved in

inflammation. These mAbs are used to treat conditions such as rheumatoid arthritis,
Crohn’s disease, psoriasis, and ankylosing spondylitis.
⬥ B-Cell Depletion: Rituximab (Rituxan), which targets CD20 on B-cells, is used in

autoimmune conditions like rheumatoid arthritis and systemic lupus erythematosus (SLE)
to deplete autoreactive B-cells.
⬥ Interleukin Inhibitors: mAbs like ustekinumab (Stelara) target interleukins (IL-12 and

IL-23) and are used to treat inflammatory diseases like psoriasis and Crohn’s disease.
C. Infectious Diseases
Monoclonal antibodies have been developed for the treatment and prevention of various
infectious diseases.
⬥ Virus Neutralization: mAbs can neutralize viruses by binding to their surface antigens,

preventing them from entering host cells. For example, casirivimab and imdevimab are
mAbs used in the treatment of COVID-19, targeting the spike protein of SARS-CoV-2 to
prevent viral entry.
⬥ Bacterial Infections: mAbs are used to neutralize bacterial toxins, such as
bezlotoxumab (Zinplava), which targets Clostridium difficile toxins to prevent
recurrence of infection.
⬥ Ebola: ZMAP, a combination of three monoclonal antibodies, was used in the treatment

of Ebola virus infection.

D. Other Diseases
Monoclonal antibodies are also used in the treatment of other conditions, such as:
⬥ Asthma and Allergies: omalizumab (Xolair) is used to treat severe asthma by targeting

immunoglobulin E (IgE), a key player in allergic reactions.

⬥ Osteoporosis: denosumab (Prolia) is used to treat osteoporosis by inhibiting osteoclast

activity, reducing bone resorption.

⬥ Cardiovascular Diseases: alirocumab (Praluent) and evolocumab (Repatha) are

PCSK9 inhibitors used to lower cholesterol levels in patients at high risk for heart
disease.
2. Diagnostic Applications of Monoclonal Antibodies
Monoclonal antibodies are widely used in laboratory diagnostics for detecting and
quantifying specific molecules, including pathogens, proteins, or other biomarkers.
A. Immunoassays: mAbs are key components of diagnostic tests that detect specific proteins,
pathogens, or biomarkers in biological samples. Common techniques include:
★ Enzyme-Linked Immunosorbent Assay (ELISA): mAbs are used in ELISA tests to

detect the presence of antigens or antibodies in blood, urine, or other fluids.

★ Western Blotting: mAbs can be used to identify proteins by their specific binding to

target antigens.
★ Lateral Flow Assays: mAbs are used in home pregnancy tests and rapid tests for

diseases like COVID-19 and influenza.

B. Immunohistochemistry: In tissue-based diagnostics, mAbs can be used to detect specific
antigens within tissue sections. This technique is crucial in cancer diagnosis, allowing
pathologists to identify the type of cancer by staining tissue samples with specific mAbs
that bind to tumor-specific markers.
C. Flow Cytometry: Monoclonal antibodies are used in flow cytometry to analyze cell
populations based on the presence of specific surface markers. This is useful for diagnosing
blood cancers like leukemia and lymphoma by detecting abnormal cell types.
D. Immunofluorescence: mAbs conjugated to fluorescent dyes are used to detect the presence
of specific molecules in cells or tissues. This technique is often used for the detection of
infections, cancer markers, and cellular localization of specific proteins.

3. Research Applications of Monoclonal Antibodies: Monoclonal antibodies are valuable tools

in biological research, providing precise and specific ways to study proteins, cells, and
biological processes.
A. Protein Detection and Quantification: mAbs are used to study proteins by detecting their
presence and measuring their concentration in biological samples. This is essential in
studying signaling pathways, gene expression, and cellular functions.
B. Cellular Marking: mAbs can be used to label specific cell types or cellular structures. This
allows researchers to isolate, track, and study individual cells in tissue culture or in vivo.
C. Targeted Delivery in Drug Development: In drug development, mAbs are used to test new
therapeutic agents. Researchers use mAbs to deliver drugs, toxins, or other agents to
specific cell types or tissues to evaluate their effectiveness in preclinical studies.
D. Model Systems: Monoclonal antibodies are often used to create animal models, such as
transgenic mice, where specific genes are knocked out or activated. mAbs can then be
used to target and study the effects of these genetic changes.
E. Vaccines and Immunotherapy Research: mAbs are also used in the development of
vaccines and immunotherapies. Researchers use mAbs to understand immune responses,
improve vaccine designs, and test new immune-modulating therapies.
 Structure and functions of major
histocompatibility complexes

Introduction to Major Histocompatibility complexes:

T and B cells both use surface molecules to recognize antigens, but they do so in different
ways. B cells, with antibodies or B-cell receptors, can directly recognize antigens on their own. T
cells, however, can only recognize antigen pieces displayed on the surface of other cells. These
pieces are held by a protein called the major histocompatibility complex (MHC), which is
encoded by the MHC genes. After a cell breaks down an antigen, the pieces are displayed on the
cell surface by the MHC molecule. The MHC acts as a vessel for presenting antigen fragments,
so T cells can recognize them with their T-cell receptors. The MHC is named for its role in
determining whether a transplanted tissue will be accepted or rejected between individuals.
Major Histocompatibility Complex (MHC) was first discovered by the immunologist Peter Gorer
in the 1930s.
The Major Histocompatibility Complex (MHC) is a genomic region found in all
vertebrates, responsible for coding proteins crucial for immune recognition. In humans, the MHC
is a gene cluster located on chromosome 6, producing proteins known as Human Leukocyte
Antigens (HLA). These MHC proteins are essential cell surface molecules that allow the
acquired immune system to identify foreign substances, which is key in determining
histocompatibility. The primary role of MHC molecules is to bind peptide antigens and present
them on the cell surface for recognition by the appropriate T-cells. Among the numerous genes
in the human MHC, those encoding class I, class II, and class III MHC proteins are particularly
important.
There are two main types of MHC molecules: class I and class II. While they have
similar final structures, they differ in how they form these structures. Class I and class II also
vary in which cells express them and the type of antigens they present. Class I MHC molecules
are found on all nucleated cells and present antigens from inside the cell, like viral proteins, to
CD8+ T cells, which kill infected cells. Class II MHC molecules are mainly found on antigen-
presenting cells (APCs) and present antigens from outside the cell, like fungi or bacteria, to
CD4+ T cells. These activated CD4+ T cells help fight extracellular infections.
 Class I and Class II MHC molecules are similar membrane proteins with related
structures and functions. Both types have been purified, and their 3D structures have been
determined through x-ray crystallography. These glycoproteins play a key role in presenting
antigens by holding peptide fragments in grooves and displaying them on the cell surface for
recognition by T cells through T-cell receptors. On the other hand, Class III MHC molecules are
unrelated to Class I and II in both structure and function, though many are involved in other
immune response processes.
Importance of MHC:
The Major Histocompatibility Complex (MHC) is a vital component of our immune
system, playing a key role in recognizing potential threats. When a foreign substance enters the
body, it must be eliminated. MHC ensures a clear distinction between the body’s essential
components and harmful invaders. MHC doesn’t act directly on foreign elements like antibodies
do. Instead, it recognizes these foreign substances and sends a signal to T lymphocytes, which
are responsible for destroying the invaders.
The primary role of MHC is to relay the correct message to the T cells, which then take
action. Depending on the situation, T cells may destroy the infected cells or eliminate just the
harmful element. Without MHC, the immune system would struggle to recognize and destroy
foreign elements, leading to serious health issues. Sometimes, certain substances that are
beneficial to the body could be mistakenly removed. In this way, MHC is crucial for signaling to
the body’s defense system, ensuring the destruction of harmful invaders while protecting
essential components.

MHC class I proteins – Structure and mode of action:

 They are encoded by the HLA-A, HLA-B, and HLA-C genes, which produce the
corresponding HLA-A, HLA-B, and HLA-C molecules.
 These class I molecules are present on nearly all nucleated cells, including platelets.
 However, they are absent in cells of the retina and brain, as well as non-nucleated red
blood cells.
 MHC Class I molecules are recognized by CD8+ co-receptors via the β2 subunit.
 These molecules sample peptides generated within the cell and communicate the cell’s
physiological condition to immune system’s effector cells, particularly CD8+ T
lymphocytes.
 Class-I MHC is a glycoprotein consisting of a 45 kDa α-chain non-covalently associated
with a 12 kDa β2-microglobulin molecule.
 The α-chain is made up of three domains: α1, α2, and α3.
 The α1 domain rests on β2-microglobulin, while the α3 domain is transmembrane,
anchoring the MHC Class I molecule to the cell membrane.
 The peptide-binding groove, located in the central region of the α1/α2 heterodimer, holds
the peptide that is being presented.
 MHC class I glycoproteins present endogenous antigens to the TCRs of CD8+ T cells.
These endogenous peptides are derived from the degradation of intracellular proteins,
including viral or tumor antigens in infected or transformed cells, through the
proteasome.
 The degradation products then move from the cytoplasm to the endoplasmic reticulum
(ER), where they are loaded onto MHC class I molecules by the peptide-loading
complex.
 CD8+ T lymphocytes express both CD8 receptors and T-cell receptors (TCRs).
 When a cytotoxic T cell's CD8 receptor binds to an MHC class I molecule and the TCR
recognizes the epitope within that molecule, the CD8+ T lymphocyte signals the target
cell to undergo apoptosis.
 This process plays a key role in cellular immunity, which is the primary defense
mechanism against intracellular pathogens like viruses and certain bacteria.

Structure of Class I and II MHC molecules

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Functions of MHC Class I:
The Major Histocompatibility Complex (MHC) Class I molecules are pivotal in the
immune system’s ability to monitor and respond to threats from intracellular pathogens, such as
viruses and some bacteria. Their functions are diverse and essential for maintaining a healthy
immune response. Important functions of MHC Class I molecules:
1. Antigen Presentation to CD8+ T Cells:
★ MHC Class I molecules are responsible for presenting endogenous antigens—peptides

derived from the degradation of intracellular proteins like viral or tumor proteins—on the
surface of cells. This allows CD8+ T cells (cytotoxic T lymphocytes) to detect and react
to cells that are infected or transformed.
★ Once a T-cell receptor (TCR) on a CD8+ T cell binds to the peptide displayed by an

MHC Class I molecule, the T cell becomes activated. This activation triggers the T cell to
carry out its immune functions, including inducing apoptosis (programmed cell death) in
infected or cancerous cells, ultimately limiting the spread of the pathogen or the growth
of abnormal cells.
2. Immune Surveillance and Response to Intracellular Pathogens:
★ MHC Class I molecules play an important role in the body’s ability to detect

intracellular pathogens. They achieve this by displaying peptide fragments of these

pathogens (like viral proteins) on the cell surface.
★ This ability is critical for the immune system to recognize and eliminate infected cells,

enabling CD8+ T cells to identify and destroy cells that harbor intracellular infections,
such as viruses or intracellular bacteria.
3. Self vs. Non-Self Recognition:
★ One of the key functions of MHC Class I molecules is to differentiate between self cells

(healthy, non-infected cells) and non-self cells (infected or abnormal cells). This
distinction is vital to prevent the immune system from mistakenly attacking the body’s
own tissues, which could lead to autoimmune diseases.
★ By presenting foreign antigens (from viruses, bacteria, or tumors) on the cell surface,

MHC Class I molecules allow the immune system to target only abnormal or infected
cells, while avoiding healthy ones.
4. Activation of Cytotoxic T Lymphocytes (CTLs):
★ The principal function of MHC Class I molecules is to activate cytotoxic T lymphocytes

(CTLs). Upon recognizing the antigen presented by MHC Class I, CD8+ T cells become
activated, multiply, and proceed to destroy target cells.
★ Cytotoxic T cells are particularly crucial in controlling viral infections and in combating

cancer cells, as they can kill cells that harbor infections or are undergoing malignant
transformation.
5. Contributing to Cellular Immunity:
★ MHC Class I molecules are integral to cellular immunity, a branch of the immune

system where T cells eliminate infected or abnormal cells directly. This is in contrast to
humoral immunity, where B cells produce antibodies that target pathogens in body
fluids.
★ Cellular immunity, and MHC Class I molecules in particular, are especially important in

defending against intracellular pathogens like viruses and some bacteria, which reside
inside host cells and are not easily targeted by antibodies alone.
6. Role in Tissue and Organ Transplantation:
★ In the context of organ transplantation, MHC Class I molecules are essential for

determining histocompatibility between the donor and recipient. This compatibility is

important because significant differences in MHC Class I molecules between the two
individuals can lead to organ rejection, as the recipient’s immune system may recognize
the transplanted organ as foreign and attack it.
7. Regulation of Immune Tolerance:
★ MHC Class I molecules also help to maintain immune tolerance—a mechanism that

prevents the immune system from attacking the body’s own healthy cells under normal
conditions. This regulation is crucial for avoiding the development of autoimmune
diseases, where the immune system wrongly targets and damages the body’s tissues.
 Antigen presentation by MHC class I molecule
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MHC class II molecules – Structure and mode of action:

 They are a type of major histocompatibility complex (MHC) typically found on antigen-
presenting cells (APCs), which are essential for initiating immune responses.
 In humans, MHC Class II proteins are encoded by the genes located in the HLA-D region
of the genome.
 MHC Class II molecules have a more restricted tissue distribution, primarily found on
macrophages, dendritic cells, B cells, and other APCs. Their expression on other cell
types, such as endothelial or epithelial cells, can be induced by IFN-γ.
 The antigens presented by MHC Class II molecules are derived from extracellular
proteins.
 MHC Class II molecules consist of β1 and β2 subunits, which allows them to be
recognized by CD4 co-receptors.
  These molecules capture the peptides from extracellular pathogens and, by interacting
with T helper cells (CD4+ T cells), and play a key role in regulating the immune
response to infections.
 MHC-II molecules are composed of dimers consisting of a 133 kDa α-chain and a 28 kDa
β-chain, which are held together by non-covalent interactions.
 Both the α-chain and β-chain consist of two domains: α1 and α2 for the α-chain, and β1
and β2 for the β-chain. These molecules are membrane-bound glycoproteins that feature
external domains, a transmembrane segment, and a cytoplasmic tail.
 The peptide-binding cleft, which is formed by the α-chain and β-chain at their proximal
ends, is an open-ended groove capable of binding antigenic peptides.
 MHC class II molecules present exogenous antigens to CD4+ T cells. Phagocytes, such
as macrophages and immature dendritic cells, engulf foreign particles via phagocytosis,
which leads to the formation of phagosomes that fuse with lysosomes. The acidic
enzymes within the lysosomes break down the engulfed proteins into various peptides.
 During the synthesis of MHC class II molecules, they are transported from the
endoplasmic reticulum (ER) through the Golgi apparatus to endosomal compartments.
 The α and β chains of MHC class II molecules are initially associated with a special
polypeptide called the invariant chain (Ii), which blocks endogenous peptides from
binding to the MHC class II binding groove.
 Once the invariant chain is removed in the acidic environment of the endosomal
compartments, peptides can bind to the groove of MHC class II molecules.
 A specific peptide, often the most immunologically dominant, is then loaded onto the
MHC class II molecules. These peptide-loaded MHC class II complexes are transported
to the cell surface, where they can be recognized by the corresponding T cell receptor
(TCR) on helper T cells for antigen presentation.

Functions of MHC Class II Molecules:

MHC (Major Histocompatibility Complex) class II molecules play a key role in the
immune system, specifically in the activation of T cells. These molecules are involved in
presenting antigens to CD4+ T cells, which are essential for initiating adaptive immune
responses. Important functions of MHC class II molecules:
1. Antigen Presentation
MHC class II molecules are primarily involved in the presentation of exogenous
antigens (i.e., antigens that are derived from outside the cell) to CD4+ T helper cells (Th cells).
These antigens are typically derived from pathogens like bacteria, viruses, or other foreign
particles that are engulfed by immune cells such as dendritic cells, macrophages, or B cells.
⬥ Phagocytosis: Antigen-presenting cells (APCs) such as dendritic cells or macrophages

engulf pathogens.
⬥ Antigen Processing: After the pathogen is engulfed, it is broken down in intracellular

vesicles, where peptides are generated from the pathogen.

⬥ Binding to MHC Class II: These antigen-derived peptides are then loaded onto MHC

class II molecules inside the vesicles. The MHC class II molecules have a peptide-
binding groove that can accommodate these peptides.
⬥ Surface Expression: The MHC class II-peptide complexes are transported to the cell

surface for presentation.

Antigen presentation by Class II MHC Molecule

Image source: https://media.geeksforgeeks.org/wp content/uploads/20221107172812/AntigenPresentation.png
2. T Cell Activation
The core function of MHC class II molecules is to activate CD4+ T cells. When an MHC
class II molecule presents a foreign peptide on the surface of an APC, a CD4+ T cell receptor
(TCR) binds specifically to this peptide-MHC class II complex.
⬥ Signal Transduction: This interaction triggers intracellular signaling in the CD4+ T cell,

leading to its activation and differentiation into different subsets of T helper cells (e.g.,
Th1, Th2, Th17, Treg) based on the cytokine environment.
⬥ Co-stimulatory Signals: In addition to the peptide-MHC interaction, other co-

stimulatory signals (e.g., CD80/86 binding to CD28 on T cells) are needed for full
activation. This ensures that the T cell response is appropriate and regulated.
3. Immune Response Regulation
Once activated, CD4+ T helper cells assist other immune cells in responding to the
infection or threat. They coordinate various aspects of the immune response by releasing
cytokines and influencing the activity of:
⬥ B cells: Th cells help B cells produce antibodies in response to pathogens.

⬥ Cytotoxic T cells (CD8+ T cells): Th cells enhance the killing ability of cytotoxic T

cells, which target and destroy infected cells.

⬥ Macrophages and dendritic cells: Th cells stimulate these cells to enhance their ability

to phagocytize pathogens and present antigens more efficiently.

4. Immune Tolerance and Autoimmunity
MHC class II molecules also play a critical role in self-tolerance by ensuring that the
immune system does not attack the body's own tissues. During development in the thymus, T
cells that recognize self-antigens presented by MHC class II molecules undergo a process of
negative selection, where self-reactive T cells are eliminated. Failure of tolerance mechanism
can lead to autoimmune diseases, where the immune system mistakenly attacks the body’s own
cells.
5. Inflammatory Response
MHC class II molecules are involved in the initiation and regulation of inflammatory
responses:
⬥ In infection or injury, they help recruit immune cells to the site of infection.
 ⬥ They influence the type of immune response based on the nature of the pathogen (e.g.,

Th1 responses for intracellular pathogens like viruses, and Th2 responses for extracellular
pathogens like parasites).
6. Antigen Specificity and Memory
⬥ MHC class II molecules help generate immunological memory by allowing CD4+ T

cells to recognize specific antigens more efficiently upon subsequent exposures.

⬥ These memory T cells enable the body to respond more rapidly and effectively to

previously encountered pathogens.

7. Involvement in Vaccination

⬥ MHC class II molecules are involved in presenting the vaccine-derived antigens to CD4+
T cells, which then help generate both humoral (antibody-mediated) and cellular
immunity.

MHC class III molecules – Structure and mode of action:

MHC class III region is involved in encoding various proteins that play important roles
in immune response, but unlike MHC class I and II, these molecules do not function in antigen
presentation. Instead, they are part of systems that regulate inflammation, immune defense, and
cellular stress responses, including:
1) Complement Proteins
MHC class III contains genes encoding several complement components, which are
crucial for innate immunity and pathogen defense. These proteins are involved in the
complement cascade, which helps eliminate pathogens by promoting inflammation,
opsonization, and lysis.
★ C2 (Serine Protease): C2 is a component of the classical complement pathway. It is

involved in the formation of the C3 convertase, which cleaves C3 to initiate the

complement cascade.
★ C4A and C4B (Pro-proteins): Both C4A and C4B are precursors of active complement

proteins. They contribute to the classical pathway of complement activation by forming

the C3 convertase, which plays a role in the opsonization of pathogens and the formation
of the membrane attack complex (MAC).
★ Factor B (Component of the Alternative Pathway): Factor B is part of the alternative

complement pathway. In this pathway, Factor B binds to C3b, and the complex activates
 the alternative C3 convertase, helping to amplify the complement response and defend
against pathogens.
2. Tumor Necrosis Factors (TNFs)
★ TNF-α (Tumor Necrosis Factor-alpha): This cytokine is involved in systemic

inflammation and is part of the body's response to infection, trauma, and cancer. It plays
a central role in immune system regulation and inflammation by inducing fever,
apoptosis, and the activation of other immune cells.
★ TNF-β (Lymphotoxin): TNF-β, also known as lymphotoxin, is closely related to TNF-α

and shares many functional similarities. It is involved in inflammatory responses and

can contribute to tissue damage and immune regulation.
3. Heat Shock Proteins (HSPs)
★ HSPs are molecular chaperones that assist in protein folding and prevent misfolded

proteins from aggregating. They are induced in response to stress (e.g., heat, oxidative
stress, infection).
★ Heat shock proteins like HSP70 and others are crucial for stabilizing newly synthesized

proteins and assisting in their proper folding.

★ They also have a role in immune system modulation by interacting with various

receptors on immune cells and influencing responses to stress and inflammation.

4. No Role in Antigen Presentation
MHC class III molecules are not involved in antigen presentation. They function more
in the regulation of the immune system and responses to infection or injury, including:
 Modulating inflammation through cytokines like TNF-α and TNF-β.

 Activating the complement system, which helps with pathogen clearance.

 Supporting cellular stress responses via heat shock proteins.

 Basic properties and functions of cytokines

Cytokines are a diverse family of small signaling proteins that play an essential role in
mediating an organism's response to injury, infection, and inflammation. These proteins function
as molecular messengers, transmitting signals between cells to regulate immune responses,
inflammation, and tissue repair processes. Cytokines are secreted in minute amounts by various
cell types, including immune cells such as macrophages, T cells, B cells, and mast cells, as well
as non-immune cells like epithelial and endothelial cells. Once secreted, cytokines exert their
effects by binding to specific receptors on target cells, initiating intracellular signaling cascades
that influence gene expression. This process may inhibit or enhance the activity of particular
genes, depending on the context, allowing cytokines to fine-tune the immune and inflammatory
responses.
Unlike endocrine hormones that typically act systemically throughout the body, most
cytokines operate in a localized manner near the cells that produce them. This ensures a targeted
and efficient response. For example, antigen-presenting cells (APCs) such as dendritic cells
release cytokines that drive T-cell activation. Upon encountering an antigen, APCs initiate a
rapid intracellular biochemical cascade, leading to the transcription of numerous genes, including
those encoding cytokines and their receptors. The activated T cells, in turn, release additional
cytokines to amplify and direct the immune response.
Most cytokines are secreted by immune system cells, while others, like type I
interferons and tumor necrosis factor-alpha (TNF-α), are produced by non-immune cells,
such as epithelial cells. These proteins play a pivotal role in regulating the development,
differentiation, and function of immune effector cells. Additionally, certain cytokines possess
direct effector functions, such as inhibiting viral replication (e.g., interferons) or inducing cell
death in tumor cells (e.g., TNF-α).
Cytokines are critical not only in regulating immune responses but also in maintaining
tissue homeostasis. Their dysregulation can lead to various pathological conditions, such as
chronic inflammation, autoimmune diseases, or cancer. For example, excessive cytokine
production may result in a "cytokine storm," a severe inflammatory state associated with
conditions like sepsis or severe viral infections. Conversely, inadequate cytokine production can
impair immunity, increasing vulnerability to infections.
 The study and therapeutic targeting of cytokines have become central to modern
medicine. Cytokine-based therapies, such as the use of monoclonal antibodies to block pro-
inflammatory cytokines like TNF-α in rheumatoid arthritis or the administration of interferons in
viral infections and cancer, highlight their importance in clinical applications.

Image source: Kuby Immunology., 7th edition

Properties of Cytokines:
Cytokines possess several distinct properties that enable them to regulate and coordinate
immune and inflammatory responses effectively. These properties define their roles as key
mediators of cellular communication in the immune system.
1. Low Molecular Weight: Cytokines are small proteins or glycoproteins with low molecular
weights, typically ranging between 8 and 30 kDa. This allows them to diffuse easily
through tissues and interact with receptors on target cells.
2. Specificity: Cytokines are highly specific in their action. They bind to specific receptors on
the surface of target cells, initiating intracellular signaling cascades. The presence of the
appropriate cytokine receptor on a cell determines whether the cell will respond to a
particular cytokine.
3. Pleiotropy: A single cytokine can exhibit multiple biological effects on different cell types.
For instance, interleukin-6 (IL-6) can promote inflammation, stimulate antibody
production, and support tissue regeneration, depending on the context and target cell.

Image source: Kuby Immunology., 7th edition

4. Redundancy: Different cytokines may have overlapping functions, meaning several cytokines
can perform similar tasks. For example, IL-1, IL-6, and TNF-α all promote inflammation.
This redundancy ensures that critical immune functions can still occur if one cytokine is
absent or deficient.

Image source: Kuby Immunology., 7th edition

5. Synergy: Cytokines often act in a synergistic manner, meaning that their combined effect is
greater than the sum of their individual effects. For instance, interferon-gamma (IFN-γ)
and TNF-α work together to enhance macrophage activation.
 Image source: Kuby Immunology., 7th edition

6. Antagonism: Some cytokines can counteract or inhibit the effects of others. For example, IL-
10 and transforming growth factor-beta (TGF-β) are anti-inflammatory cytokines that
suppress the activity of pro-inflammatory cytokines like IL-1 and TNF-α. This
antagonistic behavior helps maintain immune balance and prevent excessive responses.

Image source: Kuby Immunology., 7th edition

7. Localized Action: Most cytokines act in a localized or paracrine manner, affecting cells in
their immediate vicinity. However, in certain conditions, such as severe infections,
cytokines may act systemically, leading to widespread effects like fever or septic shock.
8. Transient Secretion: Cytokine production is highly regulated and transient. They are only
secreted in response to specific stimuli, such as infections, injury, or other immune
challenges, and their levels decrease once the stimulus is resolved. This ensures that
immune responses are appropriately timed and do not persist unnecessarily.
9. Cascade Effect: Cytokines often initiate cascades where one cytokine induces the production
of other cytokines. For example, TNF-α can stimulate the release of IL-1 and IL-6 to
amplify the immune response. This cascading property allows for rapid and coordinated
immune activation.
 Image source: Kuby Immunology., 7th edition

10. Dual Nature: Cytokines can have both beneficial and harmful effects. For instance, they are
essential for fighting infections and healing wounds, but their overproduction can lead to
inflammatory diseases or cytokine storms, as observed in severe infections like COVID-19.
11. Receptor-Mediated Action: Cytokines function by binding to specific cell surface
receptors, which triggers intracellular signaling pathways such as the JAK-STAT
pathway. These pathways lead to changes in gene expression, influencing cellular
activities like proliferation, differentiation, or apoptosis.
12. Immunoregulatory Functions: Cytokines play a vital role in regulating immune responses.
They can stimulate immune activation (pro-inflammatory cytokines like IL-1, IL-6, and
 TNF-α) or suppress immune activity (anti-inflammatory cytokines like IL-10 and TGF-β)
to maintain balance and prevent immune overreaction.
13. Short Half-Life: Cytokines are typically short-lived molecules, ensuring their effects are
limited to the time and place needed. This transient nature prevents prolonged or excessive
immune responses that could damage healthy tissues.

Types and functions of Cytokines:

Cytokines are classified into several subtypes based on their function and cell of origin:
1. Lymphokines :
★ They are protein mediators, produced by lymphocytes (typically by T cells ).
★ Can attract other immune cells, like macrophages and other lymphocytes, towards the site
of the infection and activate their immune response.
★ Lymphokines aid B cells to produce antibodies.
★ Includes Colony-stimulating factors like GM-CSF, Interferons, Interleukins, Osteoclast-
activating factor, Platelet-derived growth factor (PDGF), Transforming growth factor-
beta (tgfβ), TNFα and TNF β
Functions of lymphokines :
 Activates B cells, and inhibits macrophage function: IL-10.
 Activation of neutrophils, eosinophils, and monocyte /macrophages: GM-CSF.
 Bonemarrow– growth and differentiation: IL-3
 Bcell growth and differentiation: IL-4.
 Co-stimulator of T cells, induces growth in B cells: IL-6
 Inflammation, fever, catabolism, activation of some macrophages: TNF
 Haematopoiesis stimulators :IL-3, IL-7, GM-CSF
 Macrophage-activating activity (MAF) :INF-γ
 Proliferation of activated T and B cells: IL-2
 Inhibits T cell growth, activates macrophages:TNFβ

2. Monokines:
Produced by mononuclear phagocytic cells (e.g., macrophages), these cytokines regulate
inflammation and innate immune responses.
3. Interleukins (ILs):
Act as communicators between leukocytes, mediating a wide array of immune and
inflammatory functions. For instance, IL-1 promotes inflammation, while IL-10
suppresses it.
4. Chemokines:
Specialized cytokines that guide the migration of leukocytes to infection or injury sites,
facilitating the recruitment of immune cells during an immune response.
5. Interferons
Interferons (IFNs) are a distinct group of cytokines known for their critical role in
defending the body against viral infections, regulating immune responses, and modulating
cellular growth. They are low molecular weight proteins that are secreted primarily by cells in
response to pathogen invasion, particularly viruses, but also bacteria, parasites, and cancerous
cells. As signaling molecules, interferons act by transmitting biochemical signals between cells,
activating pathways that inhibit the replication of pathogens and orchestrate immune defense
mechanisms.

Types and functions of Interferons

Interferons are classified into three major types based on the receptors they bind to and their
function:
1. Type I Interferons:
 Includes Interferon-alpha (IFN-α) and Interferon-beta (IFN-β).

 Produced by a variety of cells, including macrophages, dendritic cells, and

fibroblasts.
 Type I interferons are primarily involved in antiviral defense. They trigger the

production of antiviral proteins within infected and neighboring cells, preventing

viral replication.
 They also enhance the activity of natural killer (NK) cells and promote antigen

presentation to adaptive immune cells.

2. Type II Interferon:
 Includes Interferon-gamma (IFN-γ).
 Produced mainly by activated T cells and NK cells.
  Type II interferon plays a crucial role in coordinating immune responses, such
as activating macrophages to kill intracellular pathogens. It is also involved in
regulating adaptive immunity by enhancing antigen presentation.
3. Type III Interferons:
 Includes Interferon-lambda (IFN-λ).

 Produced by epithelial cells and other immune cells.

 Type III interferons contribute to antiviral defense, particularly at mucosal

surfaces, and help maintain barrier immunity.

6. Tumor Necrosis Factor (TNF) as Cytokines

Tumor Necrosis Factor (TNF) refers to a group of cytokines that play a central role in
regulating immune responses, inflammation, and cell death (apoptosis). TNF is primarily
involved in controlling inflammatory processes and is a key mediator in the body's defense
against infections, cancer, and tissue injury. It is produced mainly by immune cells like
macrophages, dendritic cells, and T cells, but it can also be secreted by non-immune cells such as
fibroblasts and epithelial cells. TNF cytokines are critical for maintaining immune homeostasis
but can also contribute to pathological conditions when dysregulated.
Types of TNF
The TNF family includes several members, but the most well-known are:
1. TNF-alpha (TNF-α):
 Produced by macrophages, dendritic cells, and monocytes.

 TNF-α is heavily involved in regulating inflammation and immune responses.

 It acts on nearby cells and tissues by binding to specific receptors, TNF receptor

1 (TNFR1) and TNF receptor 2 (TNFR2), initiating complex signaling

pathways.
 TNF-α helps activate immune cells, promote the recruitment of leukocytes, and

enhance the expression of adhesion molecules on endothelial cells.

2. TNF-beta (TNF-β) (also known as lymphotoxin):
 Secreted mainly by lymphocytes.
 TNF-β has similar functions to TNF-α but is more specialized in promoting
cell-mediated immune responses and regulating lymphoid tissue development.
Functions of TNF
1. Inflammatory Regulation:
★ TNF is a key player in initiating and regulating inflammation. It induces the

production of other cytokines, such as interleukin-6 (IL-6) and interleukin-1 (IL-

1), and recruits immune cells like neutrophils and macrophages to sites of
infection or injury.
★ It increases vascular permeability, enabling immune cells to reach affected tissues

more effectively.
2. Apoptosis and Cytotoxicity:
★ TNF can induce programmed cell death (apoptosis) in infected or cancerous cells,

contributing to tumor suppression and the elimination of harmful cells.

★ TNF signaling activates caspase enzymes, which mediate apoptosis.

3. Immunomodulation:
★ TNF helps modulate adaptive immunity by enhancing antigen presentation and

activating T cells.
★ It influences the survival, proliferation, and differentiation of immune cells,

ensuring a coordinated immune response.

4. Host Defense:
★ TNF plays a crucial role in protecting the body from infections, particularly

bacterial and viral pathogens. It stimulates antimicrobial activity and the release
of reactive oxygen species (ROS) by immune cells.

Image source: Kuby Immunology., 7th edition

 Exogenous and endogenous pathways of antigen
processing and presentation
Introduction:

T cells can identify and react to protein antigens which is crucial in immune response
processes called antigen processing and presentation. Antigens undergo this process by breaking
down into peptides, attaching to Major Histocompatibility Complex (MHC) molecules, and then
being displayed on the cell membrane. The process of antigen processing starts when antigen-
presenting cells (APCs), including B cells, dendritic cells, and macrophages, internalise the
protein antigens. Antigen processing involves the enzymatic breakdown of the antigens into
smaller peptide fragments inside these cells.
A peptide-MHC complex is formed when the antigens are converted into peptides and
then bind to MHC molecules in the cytoplasm of the cell. The cytoplasm of the cell processes the
peptides produced from endogenous antigens in the case of MHC Class I molecules. Tumour,
viral, bacterial, and cellular proteins are examples of these endogenous antigens. These peptides
are bound by Class I MHC molecules, which then display them on the cell surface. This process
is known as cytosolic pathway.
However, peptides from external antigens that are absorbed by the cell through
phagocytosis or endocytosis are bound by MHC Class II molecules. Extracellular bacteria,
viruses, and other foreign particles are examples of these exogenous antigens. The endocytic
pathway processes the antigens and breaks them down into peptides. The resultant peptides
attach to Class II MHC molecules and travel to the cell membrane, where CD4+ T helper cells
can encounter and recognise them. This process is called as endocytic pathway.
MHC Class I and Class II molecules are essential for T cell activation because they
deliver antigen-derived peptides. The key to trigger an immune response to particular antigen is
the interaction between the peptide-MHC complex on antigen-presenting cells and the T cell
receptor on T lymphocytes. T lymphocyte activation and proliferation, as well as the
coordination of many immunological effector mechanisms, are the results of a series of immune
reactions set off by this recognition.
This mechanism is essential to immune surveillance, infection prevention, and the
removal of aberrant or contaminated cells. Target cells are typically cells that have contracted an
intracellular pathogen, such as a virus. Targets may also include changed self-cells, such as
cancerous cells, ageing body cells, or allogeneic cells from a graft. It is also a basic component
of the adaptive immune response.

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MHC class I Pathway & MHC Class II Pathway

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Pathways for Processing and Presentation:
The immune system has two distinct processes to get rid of external and intracellular
antigens. Exogenous antigens, which are produced inside the cell, are processed in the endocytic
pathway and displayed on the membrane with class II MHC molecules, while endogenous
antigens, which are taken up by endocytosis, are processed in the cytosolic system.
In eukaryotic cells, protein levels are strictly controlled. Every protein experiences
continuous turnover and is broken down at a rate known as its halflife. Certain proteins, such as
cyclins, transcription factors, and vital metabolic enzymes, have incredibly short half-lives, while
proteins that are denatured, misfolded, or otherwise abnormal are also quickly eliminated.
Endogenous Antigens: The Pathway of Cytoplasm
The same pathways used in the regular turnover of intracellular proteins are also used in
the degradation of endogenous antigens for presentation with class I MHC molecules.
1. Peptides for Presentation Are Generated by Protease Complexes Called Proteasomes:
Protease Complexes Known as Proteasomes generate Peptides for Presentation. All cells
have a cytosolic proteolytic mechanism that breaks down intracellular proteins into small
peptides. A tiny protein known as ubiquitin is frequently linked to the proteins that are the targets
of proteolysis. A multipurpose protease complex known as a proteasome is capable of breaking
down ubiquitin-protein conjugates. Peptide bonds between two or three distinct amino acid
combinations can be broken by a proteasome in an ATP-dependent manner.
2. Peptides Are Transported from the Cytosol to the Rough Endoplasmic Reticulum:
The hydrolysis of ATP is necessary for TAP to transfer peptides produced by the
proteasome in the cytosol into the RER. Two proteins, TAP1 and TAP2, form the membrane-
spanning heterodimer known as the transporter protein, or TAP (transporter associated with
antigen processing). The TAP1 and TAP2 proteins each feature a domain that extends into the
RER lumen and an ATP-binding domain that extends into the cytosol in addition to their many
transmembrane regions. Both TAP1 and TAP2 are members of the ATP-binding cassette protein
family, which is present in the membranes of many bacteria and other cells. These proteins
facilitate the ATP-dependent transport of peptides, sugars, ions, and amino acids. The ideal
peptide length for class I MHC binding is 8–10 amino acids, for TAP’s best affinity.
Furthermore, TAP seems to favour peptides containing the recommended anchor residues for
class I MHC molecules, which are hydrophobic or basic carboxyl-terminal amino acids. TAP is
therefore designed to carry peptides that will engage with class I MHC molecules.
3. Peptides Assemble with Class I MHC Aided by Chaperone Molecules:
The class I MHC molecule's α chain and β2-microglobulin components are made on
polysomes throughout the rough endoplasmic reticulum, just like other proteins. A peptide in the
class I molecule's binding groove is necessary for these elements to assemble into a stable class I
MHC molecular complex that can leave the RER. The folding of polypeptides is facilitated by
molecular chaperones, which participate in the multi-step assembly process. Calnexin, an
endoplasmic reticulum resident membrane protein, is the first molecular chaperone implicated in
class I MHC assembly. Calnexin encourages the folding of the free class I α chain by associating
with it. Calnexin is released when β2-microglobulin attaches to the α chain, and the class I
molecule binds to tapasin and the chaperone calreticulin. The class I molecule is brought close to
the TAP transporter by the tapasin (TAP-associated protein), which enables it to take up an
antigenic peptide. The class I molecule has enhanced stability as a result of peptide binding, and
it can separate from tapasin and calreticulin, leave the RER, and travel through the Golgi to the
cell surface. Calnexin and calreticulin complexes have been found to be associated with another
chaperone protein, ERp57.

MHC I Endogenous Antigen-Processing Pathway

Image source: https://www.bio.davidson.edu/movies/Immunology/Students/spring2006/Witcher/TAP.html
Exogenous antigen processing pathway:
The exogenous antigen processing pathway refers to the way the immune system
processes and presents antigens that originate from outside the cell, typically from pathogens like
bacteria, viruses, and other environmental agents. This process is crucial for activating the
immune response, particularly in the context of CD4+ T cells. It involves the following steps:
Step 1: Antigen Uptake (Endocytosis and Phagocytosis)
Exogenous antigens are those derived from outside the cell, such as pathogens (bacteria,
fungi, or viruses) or other foreign particles. These antigens are taken up by antigen-presenting
cells (APCs), including dendritic cells, macrophages, and B cells, through two primary
mechanisms:
⬥ Phagocytosis: This is the process by which larger particles such as whole bacteria are
engulfed by the antigen-presenting cell. The antigen is enclosed within a phagosome, a
membrane-bound vesicle.
⬥ Endocytosis: This process involves the internalization of smaller antigens, including
soluble antigens or viral particles, into the cell through the invagination of the plasma
membrane.
Step 2: Antigen Processing (Degradation)
Once the antigen is inside the cell, it must be broken down into smaller fragments, which
will later be presented to T cells. The antigen is typically enclosed in an acidic vesicle known as
the phagosome or endosome. The phagosome or endosome fuses with lysosomes, which contain
a variety of hydrolytic enzymes (such as proteases) that degrade the antigen into small peptide
fragments. These peptides are generally between 8-20 amino acids long, although the length
may vary slightly depending on the processing. The degradation of the antigen may occur in
several stages:
1 Initial processing: Antigens are broken down in an acidic environment within the
vesicle by enzymes like cathepsins.
2 Peptide generation: These fragments are generated as the antigen is processed
into small peptides suitable for binding to MHC class II molecules.
Step 3: Loading of Peptides onto MHC Class II Molecules
After the antigen has been processed into peptides, these peptides must be loaded onto
MHC class II molecules to be presented to CD4+ T cells. MHC class II molecules are
synthesized in the endoplasmic reticulum (ER), where they are assembled and associate with
an invariant chain (Ii). The invariant chain prevents premature binding of peptides to the MHC
II molecule before the appropriate antigen peptides are available. Once the MHC II molecules
leave the ER, they are transported to the endosomal compartment (which contains the
processed peptide fragments). In the endosome, the invariant chain is degraded by proteases,
leaving a small fragment called the CLIP (Class II-associated invariant chain peptide) bound to
the MHC II peptide-binding groove. The CLIP is exchanged for the antigenic peptides via the
action of the HLA-DM molecule (in humans) or DM (in mice). HLA-DM facilitates the removal
of CLIP and the binding of the processed antigen peptides to the MHC II molecule.
Step 4: Transport of MHC II-Peptide Complex to the Cell Surface
Once the antigenic peptides have been successfully loaded onto MHC class II molecules,
the MHC II-peptide complex is transported to the cell surface. This occurs through vesicular
trafficking within the antigen-presenting cell. MHC II molecules, now bound to their antigenic
peptides, are packaged into vesicles that move toward the cell membrane. The vesicles fuse with
the plasma membrane, presenting the MHC II-peptide complex on the surface of the cell.
Step 5: Recognition by CD4+ T Cells
The MHC II-peptide complex on the surface of the antigen-presenting cell can now be
recognized by CD4+ T cells. The recognition occurs through the T cell receptor (TCR), which
is specific for a particular peptide-MHC complex. The TCR on CD4+ T cells specifically
recognizes the antigenic peptide bound to the MHC II molecule, and this interaction is stabilized
by co-receptors such as CD4, which enhances the binding affinity between the TCR and the
MHC II molecule. Additionally, co-stimulatory molecules (e.g., CD80/86 on the APC and CD28
on the T cell) provide essential secondary signals required for T cell activation.
Step 6: T Cell Activation and Differentiation
Upon successful recognition and binding of the TCR to the peptide-MHC II complex, the
CD4+ T cell becomes activated. Activation of T cells involves several intracellular signaling
events like TCR signaling, Co-stimulation molecules like CD28 and CD80/86 is essential for
full T cell activation. The activated T cell undergoes clonal expansion and differentiation into
various subsets of helper T cells (such as Th1, Th2, Th17, or Tfh) depending on the cytokine
environment. These helper T cells then perform a variety of function like the activation of
macrophages and the production of pro-inflammatory cytokines (e.g., IFN-γ), which help in the
defense against intracellular pathogens, support B cell differentiation and antibody production,
particularly in response to extracellular pathogens like helminthes, Play a role in mucosal
immunity and in the defense against fungal infections.

MHC II Exogenous Antigen-Processing Pathway

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Comparision of exogenous and endogenous pathways of antigen processing

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 Gell and Coombs’ ClassifiCation and various Types
of Hypersensitivities
Introduction:
Introduced by Robert Gell and Philip Coombs in 1963, this classification became a cornerstone
for immunological research and clinical diagnosis. It was revolutionary in systematically categorizing
hypersensitivity reactions, which are exaggerated or misdirected immune responses that harm the body.
This system helped bridge the gap between the physiological mechanisms of the immune system and
their pathological consequences.
The Gell and Coombs classification provides a clear framework for comprehending how the
immune system reacts under normal and pathological circumstances. It highlights the diverse ways the
immune system can malfunction either by overreacting to harmless substances (as seen in allergies) or
misidentifying self-antigens as threats (leading to autoimmune diseases). This categorization helps
immunologists and clinicians to systematically study immune responses, paving the way for targeted
interventions. By breaking down hypersensitivity into distinct mechanisms (Types I–IV), researchers
can better understand the underlying processes, such as antibody-mediated damage or T-cell-driven
inflammation. Significance of classification is as follows:
Improved Diagnosis:
One of the most significant contributions of this classification lies in its diagnostic utility.
Specific symptoms can be linked to corresponding immune mechanisms, ensuring precision in
identifying the type of hypersensitivity reaction. For instance:
 Type I hypersensitivity symptoms like immediate swelling and itching indicate an IgE-

mediated reaction.
 Type IV hypersensitivity, characterized by delayed redness and inflammation, points toward a

T-cell-mediated response.
By narrowing down the root cause of symptoms, healthcare providers can offer faster and more
accurate diagnoses, reducing the risk of misdiagnosis and enabling effective management of conditions
ranging from food allergies to autoimmune disorders.
Tailored Treatment:
The classification system guides the development and administration of treatments that are
specific to the type of immune response involved as mentioned below
 For Type I hypersensitivity, antihistamines, corticosteroids, or epinephrine are used to

counteract mast cell and basophil activation.

  For Type II reactions, treatments might include immune suppression or plasma exchange to

reduce antibody levels.

 Type III responses may require interventions to manage immune complex deposition, such as

anti-inflammatory drugs or immunosuppressive agents.

 In Type IV hypersensitivity, therapies often focus on controlling T-cell activity using

immunosuppressive drugs like corticosteroids or monoclonal antibodies.

This targeted approach minimizes side effects while maximizing efficacy, ensuring that
patients receive treatments tailored to the specific immune mechanisms involved in their condition.
Promoting Research
The Gell and Coombs classification has been instrumental in advancing immunological and
medical research. By providing a framework to study hypersensitivity mechanisms, it has inspired the
exploration of complex immune processes and the development of innovative treatments. For example:
 Autoimmune disorders: Insights from this classification have fueled breakthroughs in

understanding diseases like systemic lupus erythematosus (SLE) and type 1 diabetes.
 Allergy vaccines: Research into Type I hypersensitivity has supported efforts to create

therapies that reduce allergic reactions.

 Drug discovery: The classification aids pharmaceutical companies in designing drugs that

specifically target components of hypersensitivity reactions, like cytokine inhibitors for Type IV
responses.

Type I Hypersensitivity

Type I hypersensitivity, often referred to as immediate hypersensitivity or allergic reaction, is a

type of immune response that occurs quickly, usually within minutes, after exposure to an allergen.
This hypersensitivity is mediated by IgE antibodies, which play a key role in driving the reaction.
Allergic conditions such as hay fever, asthma, hives, and anaphylaxis are manifestations of Type 1
hypersensitivity. It is triggered when the immune system mistakenly identifies harmless substances like
Environmental: Pollen, dust mites, pet dander.
★ Foods: Peanuts, shellfish, eggs, dairy.

★ Insect Venoms: Bee or wasp stings.

★ Medications: Penicillin, aspirin, or other drugs.

Mechanism of Type I Hypersensitivity: This involves 3 important phases
1. Sensitization Phase:
 On the first exposure to an allergen (e.g., pollen, dust mites, or food proteins), antigen-

presenting cells (APCs) like dendritic cells capture and process the allergen.
 These cells present the allergen to naïve helper T-cells (Th2), prompting their activation.

 Th2 cells release cytokines like interleukin-4 (IL-4) and interleukin-13 (IL-13), which

stimulate B-cells to produce allergen-specific IgE antibodies.

 The IgE antibodies bind to high-affinity receptors (FcεRI) on mast cells and basophils,

"arming" these cells for future reactions.

2. Reaction Phase:
 On subsequent exposures, the allergen cross-links IgE molecules on the surface of

sensitized mast cells and basophils.

 This cross-linking triggers the release of granules containing histamine, leukotrienes,

prostaglandins, and other inflammatory mediators.

3. Effects of Mediators: These effects lead to the rapid onset of symptoms, typically within
minutes.
 Histamine: Causes vasodilation, increased vascular permeability, bronchoconstriction,

and itching.
 Leukotrienes and Prostaglandins: Enhance inflammation, prolong smooth muscle

contraction, and increase mucus production.

Second exposure

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 Type I hypersensitivity can range from mild allergic reactions to life-threatening emergencies.
Common examples include:
1. Local Reactions:
Allergic Rhinitis (Hay Fever): Symptoms include sneezing, nasal congestion, watery
eyes, and itching.
Asthma: Characterized by wheezing, shortness of breath, and chest tightness due to
airway inflammation and constriction.
Atopic Dermatitis (Eczema): Chronic skin inflammation, often linked to Type 1
hypersensitivity.
2. Systemic Reactions:
Urticaria (Hives): Raised, red, and itchy welts on the skin.
Angioedema: Swelling of deeper layers of the skin, often around the eyes, lips, or
throat.
Anaphylaxis: The most severe form, marked by widespread vasodilation, airway
constriction, and shock. It requires immediate treatment with epinephrine.

Cells and molecules involved in this mechanism:

✿ Role of Mast Cells: Mast cells play a central role as they are stationed in tissues (skin,

respiratory, and gastrointestinal tracts) where allergen exposure is likely. When IgE binds to
mast cells, it primes them for explosive degranulation upon allergen exposure.
✿ Basophils: These are circulating cells analogous to mast cells. They amplify the allergic

response by releasing inflammatory mediators during the reaction phase.

✿ Cytokines: IL-4 promotes IgE synthesis, while IL-5 recruits eosinophils, contributing to tissue

damage and inflammation.

✿ Eosinophils: These immune cells are recruited to the site of inflammation, exacerbating tissue

damage via release of toxic granules like eosinophilic cationic protein.

Histological changes observed:
★ Edema: Tissue swelling due to leakage of fluids from dilated blood vessels.

★ Epithelial Damage: Prolonged exposure can lead to destruction of epithelial cells, particularly

in chronic asthma or dermatitis.

 ★ Inflammatory Cell Infiltration: A flood of immune cells (eosinophils, neutrophils, etc.) at the

site of allergen exposure.

Management of Type 1 Hypersensitivity:

1. Avoiding exposure to known allergens.
2. Pharmacological Interventions:
 Antihistamines: Block histamine receptors to alleviate itching and swelling.

 Corticosteroids: Reduce inflammation in severe cases.

 Leukotriene Receptor Antagonists: Help manage asthma and allergic rhinitis.

 Epinephrine: Used as an emergency treatment for anaphylaxis.

3. Immunotherapy:
 Allergen Desensitization: Gradual exposure to increasing amounts of the allergen to

build tolerance over time.

Type II Hypersensitivity

Type II hypersensitivity, often termed cytotoxic hypersensitivity, is an immune reaction in

which the body's defense system mistakenly targets its own cells for destruction. It is primarily
mediated by IgG or IgM antibodies, which bind to antigens located on cell surfaces. On a cellular level,
Type 2 hypersensitivity causes tissue damage. It leads to Complement activation and immune cell
recruitment can damage blood vessels, release of cytokines and chemokines amplifies local
inflammation, worsening tissue injury, lysis and phagocytosis resulting in loss of functional cells. The
hallmark of this hypersensitivity is the direct attack on cells, leading to their destruction through
various mechanisms.
Mechanism of Type 2 Hypersensitivity:
Type 2 hypersensitivity involves the interaction of antibodies with surface antigens on host
cells. This triggers immune-mediated destruction via the following processes:
1. Complement Activation: a pivotal process in Type 2 hypersensitivity:
When IgG or IgM antibodies bind to antigens expressed on the surface of target cells, they
activate the complement system, a group of proteins circulating in the blood.
 The complement cascade ultimately forms the Membrane Attack Complex (MAC), a pore-
like structure that disrupts the cell membrane, leading to osmotic imbalance and lysis (bursting)
of the cell.
Complement proteins like C3a and C5a also act as chemoattractants, drawing immune cells
(e.g., neutrophils and macrophages) to the site of injury and promoting inflammation.

2. Antibody-Dependent Cellular Cytotoxicity (ADCC): This mechanism involves:

Natural Killer (NK) cells, which recognize and bind to the Fc region of antibodies coating the
surface of the target cell.
Once bound, NK cells release cytotoxic molecules like perforin (which forms pores in the target
cell membrane) and granzymes (which trigger apoptosis, or programmed cell death).
This process is critical for removing infected or abnormal cells.
3. Opsonization and Phagocytosis
Opsonization is the "tagging" of target cells by antibodies. These antibodies act like flags,
signaling phagocytic cells such as macrophages and neutrophils to engulf and digest the
tagged cells.
Phagocytes recognize the antibodies through their Fc receptors and proceed to engulf the cells
into a structure called the phagosome, where the cell is broken down by enzymes.

Type II Hypersensitivity
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Examples:
Autoimmune Diseases
1. Autoimmune Hemolytic Anemia (AIHA):
 IgG or IgM antibodies target antigens on red blood cell (RBC) membranes. This leads to

complement activation and phagocytosis of RBCs by macrophages in the spleen and

liver.
 Symptoms include fatigue, jaundice, shortness of breath, and dark-colored urine due to

excessive hemoglobin breakdown.

 Autoimmune disorders, certain infections, or medications.

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2. Goodpasture Syndrome:
 IgG antibodies bind to the basement membrane in the glomeruli of the kidneys and

alveoli of the lungs. This causes complement activation and inflammation.

  Hematuria (blood in urine), proteinuria, respiratory symptoms (e.g., coughing blood),

and kidney failure.

 Affects both renal and pulmonary systems and often progresses rapidly.

3. ABO incompatibility:
 ABO incompatibility is a classic example of Type 2 hypersensitivity, which is an
antibody-mediated cytotoxic reaction. This occurs when a person receives a blood
transfusion with an incompatible ABO blood type, leading to the destruction of the
transfused red blood cells (RBCs) by the recipient's immune system.
 The binding of IgM antibodies to the incompatible RBCs activates the complement
system.
 This leads to the formation of the Membrane Attack Complex (MAC), which lyses the
transfused RBCs.
 Complement activation releases inflammatory mediators like C3a and C5a, which
recruit immune cells and amplify the inflammatory response.
 The destruction of RBCs results in the release of hemoglobin into the bloodstream,
which can lead to complications like jaundice or kidney damage.

Type II Hypersensitivity - ABO Incompatibility

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4. Myasthenia Gravis:
 Autoantibodies block acetylcholine receptors (AChRs) at the neuromuscular
junction, preventing the transmission of nerve signals to muscles.
 Muscle weakness that worsens with activity (e.g., difficulty speaking, swallowing, or
breathing in severe cases).
 Affects voluntary muscles, especially those controlling the eyes, face, and throat.

Type II Hypersensitivity reaction, Myasthenia gravis mechanism

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3. Graves’ Disease:
 Autoantibodies mimic thyroid-stimulating hormone (TSH) by binding to TSH receptors

on the thyroid gland. This leads to excessive thyroid hormone production

(hyperthyroidism).
 Weight loss, rapid heartbeat, sweating, tremors, and protrusion of the eyes
(exophthalmos).

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Type III Hypersensitivity
Type III hypersensitivity refers to an immune system dysfunction where immune complexes are
composed of antigens bound to antibodies and are formed in excess and fail to be efficiently cleared
from the circulation. Instead, these complexes deposit in various tissues, initiating inflammatory
processes that can lead to tissue damage and organ dysfunction. It typically involves soluble antigens
rather than cell-bound ones, differentiating it from Type II hypersensitivity. Type III hypersensitivity,
also known as immune complex-mediated hypersensitivity.

Mechanism of Type III Hypersensitivity (Immune Complex-Mediated):

1. Immune Complex Formation: Antigens involved are typically soluble (not cell-bound), and can be
Exogenous, e.g., microbial antigens from infections (like streptococci or hepatitis B) and
Endogenous, e.g., self-antigens in autoimmune conditions (like nuclear antigens in SLE).
Antibodies, mainly IgG and sometimes IgM, bind these soluble antigens in the circulation.
Small or intermediate-sized complexes are poorly cleared by the mononuclear phagocyte system
and tend to deposit in tissues. Large complexes are usually cleared efficiently by the liver and
spleen. Factores like Antigen excess (relative to antibody)., deficiencies in complement or
phagocytic clearance mechanisms, continuous presence of antigen, e.g., in chronic infections or
autoimmunity lead to type III hypersensitivity.
2. Deposition in Tissues: Immune complexes deposit in areas with high filtration rates, such as
capillaries, glomeruli of the kidneys, synovial joints, and alveoli in the lungs. Factors
influencing deposition include antigen concentration, antibody affinity, and blood flow
dynamics.
3. Activation of the Complement System: Immune complexes trigger the complement cascade,
particularly the generation of C3a and C5a, which are potent inflammatory mediators. This
complement activation enhances the recruitment of immune cells to the site of deposition.
4. Inflammation and Tissue Damage: Neutrophils and macrophages are recruited by the complement
components and release proteolytic enzymes and reactive oxygen species (ROS). This
inflammatory response damages surrounding tissues, leading to localized or systemic effects.
For example, deposits in the kidneys cause glomerulonephritis, impairing filtration and urine
production, leads to chronic inflammation and reduced mobility, as seen in rheumatoid arthritis
and Immune complexes in blood vessel walls lead to vasculitis (inflammation of blood vessels).
 Persistent immune complex deposition can cause prolonged inflammation, resulting in fibrosis
and organ damage over time.

Type III
hypersensitivity

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Examples of Type III Hypersensitivity:

1. Systemic Lupus Erythematosus (SLE): Immune complexes form between autoantibodies and
nuclear antigens (e.g., DNA). These deposits cause inflammation in the kidneys
 (glomerulonephritis), joints (arthritis), and skin (malar rash). Complement levels (C3 and C4)
are often depleted due to activation.
2. Post-Streptococcal Glomerulonephritis: Immune complexes are formed against streptococcal
antigens following an infection. Deposits in the kidney's glomeruli lead to hematuria (blood in
urine), proteinuria, and kidney dysfunction.
3. Rheumatoid Arthritis: Immune complexes deposit in the synovial joints, causing chronic
inflammation, cartilage degradation, and joint deformities.
4. Serum Sickness: Occurs after exposure to foreign proteins such as anti-sera or drugs, leading to
immune complex formation. Symptoms include fever, rash, swollen lymph nodes, and joint
pain.
5. Hypersensitivity Pneumonitis: Chronic exposure to inhaled antigens (e.g., mold or bird
proteins) leads to immune complex deposition in the lungs, causing inflammation and fibrosis.
6. Arthus Reaction: A localized reaction occurs after repeated exposure to an antigen, leading to
immune complex formation in the skin. Symptoms include redness, swelling, and necrosis at the
injection site (common in vaccine sensitivity).

Type IV Hypersensitivity:
Type IV hypersensitivity, also known as delayed-type hypersensitivity (DTH), is a
cell-mediated immune response that takes several hours to days to develop after exposure to
an antigen. Unlike Types I-III hypersensitivity, which are mediated by antibodies, Type IV
hypersensitivity is entirely dependent on T-cells and their interaction with antigens. The
delayed nature of this hypersensitivity is a hallmark feature, as the immune response peaks
between 24 to 72 hours after antigen exposure.

Mechanism of Type IV hypersensitivity:

1. Antigen Presentation: The process starts when an antigen-presenting cell (APC)—such as a
macrophage, dendritic cell, or Langerhans cell—captures and processes the antigen. The
antigen is presented to naïve T-cells on a major histocompatibility complex (MHC) molecule:
MHC Class II is used to present the antigen to CD4+ Helper T-cells and MHC Class I is used
to present the antigen to CD8+ Cytotoxic T-cells. The interaction between the APC and the T-
cell requires costimulatory signals, such as those provided by CD80/CD86 molecules on the
 APC, binding to CD28 on the T-cell. This activates naïve T-cells to differentiate into effector T-
cells, which mediate the immune response.
2. T-cell Activation: Helper T-cells (Th1 subset) play a dominant role in Type IV hypersensitivity.
Upon activation, Th1 cells release cytokines such as Interferon-gamma (IFN-γ) that activates
macrophages and amplifies their ability to engulf pathogens. Tumor Necrosis Factor-alpha
(TNF-α) which promotes inflammation and vascular permeability. Interleukin-2 (IL-2) which
drives the proliferation and activation of other T-cells and immune cells. Cytotoxic T-cells
(CD8+ T-cells) also contribute by directly killing infected or damaged host cells where CD8+ T-
cells release perforins, which form pores in the membrane of the target cells. They also release
granzymes, which enter through these pores to induce apoptosis (programmed cell death).
3. Recruitment of Immune Cells: The cytokines secreted by activated T-cells recruit various
inflammatory cells to the site, like Macrophages which are heavily activated by IFN-γ and
become "hyperactivated," releasing enzymes like lysosomal hydrolases and matrix
metalloproteinases that degrade tissues. Neutrophils are recruited by chemokines to amplify the
inflammatory response. Fibroblasts in chronic cases, are activated to contribute to tissue repair
but may also lead to fibrosis. As these immune cells accumulate at the site, they release
inflammatory mediators such as Reactive Oxygen Species (ROS), Nitric Oxide (NO) and
Proteolytic Enzymes, which destroy pathogens but also damage host tissues.
4. Tissue Destruction occurs due to the release of Proteolytic enzymes from macrophages and
Cytotoxic molecules from CD8+ T-cells. Inflammation is amplified by cytokine signaling loops,
creating a vicious cycle of tissue damage and immune activation.
5. Delayed Response: Unlike other types of hypersensitivity, Type IV hypersensitivity requires
time for T-cells to be activated and recruited to the site of antigen exposure. This results in a peak
response at 24–72 hours after exposure, explaining the "delayed" nature of the hypersensitivity.
The reaction persists as long as the antigen is present and the immune cells remain activated,
which can lead to chronic inflammation in some cases.
Examples of Type IV Hypersensitivity
★ Tuberculin Reaction: In the Mantoux test for tuberculosis, injected PPD antigen causes a

localized swelling and redness mediated by Th1 cells and macrophages.

★ Contact Dermatitis: Exposure to allergens like nickel or poison ivy activates Th1 cells, leading

to redness, itching, and vesicle formation.

 ★ Granuloma Formation: Chronic antigen exposure in diseases like tuberculosis leads to

granulomas—clusters of macrophages and Th1 cells surrounded by fibrous tissue.

Type IV Hypersensitivity
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 Introduction to concepts of autoimmunity,
Primary and secondary Immunodeficiencies
All individuals have some level of immune recognition of their own tissues. In fact, for
T-cells to mature in the thymus, they must be positively selected based on their ability to
recognize the body’s own MHC molecules. Additionally, in most people, except those with
severe immune deficiencies self-reactive B-cells, T-cells that recognize self-peptides presented
by self-MHC, and autoantibodies (antibodies that react with the body’s own components) can be
found in the bloodstream. In people without autoimmune diseases, these autoantibodies are
mostly low-affinity IgM types, commonly produced by CD5+ B-1 cells as part of the body’s
natural antibody defense.
The term autoimmune disease is used when this self-reactivity leads to tissue damage or
illness. Some level of harmless autoimmunity may play a beneficial role in clearing out old or
damaged cells and molecules. Therefore, a mild degree of autoimmunity is considered normal
and usually doesn't cause disease. During lymphocyte development in the central lymphoid
organs, gene rearrangements naturally lead to the creation of some lymphocytes that can
recognize the body’s own antigens. Normally, these self-reactive cells are either eliminated or
kept under control through various regulatory mechanisms. These processes establish self-
tolerance, ensuring that the immune system does not target the body’s own tissues. Self-
tolerance relies on the coordinated function of multiple mechanisms that act at various stages and
locations during immune system development. The immune system uses different strategies to
prevent the activation of self-reactive lymphocytes and to avoid damage to the body's own
tissues. Autoimmune diseases occur only when immune tolerance fails and harmful self-reactive
lymphocytes are not properly controlled or eliminated.
The concept of autoimmunity was first introduced in the early 20th century by Paul
Ehrlich, who referred to it as "horror autotoxicus." Autoimmune responses are similar to normal
immune reactions against pathogens, but instead of targeting foreign invaders, they are triggered
by the body’s own antigens—called autoantigens. These responses lead to the production of self-
reactive immune cells and antibodies, known as autoantibodies, which attack the body’s own
tissues. When these self-directed responses become uncontrolled, they can result in a range of
chronic conditions known as autoimmune diseases. The breakdown can occur through several
mechanisms, which are grouped into central, peripheral, genetic, and environmental factors.
These diseases vary widely in terms of how severe they are, which tissues are affected, and the
immune mechanisms responsible for the resulting tissue damage.
Causes of Autoimmunity:
1. Failure of Central Tolerance
Central tolerance takes place during early lymphocyte development in the thymus for T-
cells and the bone marrow for B-cells. Its primary role is to eliminate strongly self-reactive
lymphocytes before they can enter the circulation. T-cells in the thymus undergo positive
selection (for recognizing self-MHC) and negative selection (elimination if they bind too
strongly to self-antigens). B-cells in the bone marrow that bind to self-antigens are either
eliminated by apoptosis (clonal deletion) or edited via receptor editing (a new light chain gene
rearrangement) or made anergic (unresponsive).
2. Failure of Peripheral Tolerance
Even after central tolerance, some autoreactive cells escape into the periphery. Peripheral
tolerance mechanisms which act as a second line of defense to prevent autoimmunity are:
a. Anergy (Functional Inactivation): Autoreactive lymphocytes that recognize self-antigen
without the necessary co-stimulatory signals (e.g., CD28 binding to B7) become anergic or
functionally inactive. In inflammation, antigen-presenting cells (APCs) may provide co-
stimulation to autoreactive cells, activating them.
b. Regulatory T-cell (Treg) Suppression: Tregs, expressing FOXP3, suppress immune
responses by releasing IL-10 and TGF-β (anti-inflammatory cytokines), direct cell–cell contact
inhibition, consumption of IL-2 to stop effector T-cells from receiving the growth signals.
Defects in Treg development/function lead to loss of immune suppression.
c. Activation-Induced Cell Death (AICD): Chronic activation of self-reactive T-cells induces
apoptosis via Fas-FasL interactions. Defects in Fas pathway prevent this cell death, allowing
autoreactive cells to persist.
3. Molecular Mimicry:
Some pathogens contain antigens structurally similar to self-antigens. Immune response
against the pathogen may accidentally cross-react with self-tissue due to similar epitopes. For
example: Streptococcal M protein resembles cardiac myosin. Antibodies formed against the M
protein attack heart tissue which leads to Rheumatic fever
4. Epitope Spreading:
Initial tissue damage or immune response exposes hidden or cryptic epitopes which were
not previously exposed to the immune system. This leads to a broader immune response against
multiple self-antigens in the same or nearby tissues. For example: In Systemic Lupus
Erythematosus (SLE), Initial response may target nuclear proteins, later it spread to DNA,
histones, etc.
5. Neoantigen Formation:
Self-molecules can be altered chemically or structurally by drugs, infections, oxidative
stress. These modifications generate neoantigens i.e., newly formed self-antigens that now
appear foreign to the immune system.
6. Genetic Predisposition:
Genetics play a major role in susceptibility to autoimmunity. Many autoimmune diseases
are associated with specific HLA alleles, especially MHC Class II molecules involved in antigen
presentation. Some genetic factors affects co-stimulation and Treg function, modify T-cell
receptor signaling, affect T-cell survival and expansion.
7. Environmental factors often serve as critical initiators or amplifiers of autoimmune
responses, particularly in individuals with an underlying genetic susceptibility. These external
influences can disrupt immune homeostasis, promote inflammation, or modify self-antigens in
ways that break tolerance and trigger autoimmunity.
Some of the important factors are:
★ Infections (Viral and Bacterial): Pathogens can provoke autoimmunity leading the

immune system to mistakenly attack self-tissues.

★ Toxins and Chemicals: Environmental toxins, including certain drugs and industrial

chemicals, may modify self-proteins to create neoantigens that the immune system no
longer recognizes as self.
★ Smoking: A well-established risk factor for diseases like rheumatoid arthritis and

systemic lupus erythematosus, smoking promotes chronic inflammation and impaired

immune regulation.
★ Ultraviolet (UV) Radiation: UV exposure, particularly in genetically predisposed

individuals, can lead to keratinocyte apoptosis, releasing nuclear antigens and triggering
or worsening cutaneous lupus erythematosus and systemic lupus.
 ★ Diet and Gut Microbiota: The composition of the gut microbiome and dietary patterns

can modulate immune function by influencing T-regulatory cell development, intestinal

permeability, and systemic inflammation factors increasingly recognized in autoimmune
conditions such as inflammatory bowel disease and type 1 diabetes.
Types of Autoimmunity:
There are two main types of autoimmune diseases based on their mechanisms namely the
organ specific and systemic autoimmune diseases.
A. Organ-Specific Autoimmunity:

★ mediated by direct cellular damage in which the immune system targets and destroys
specific cells, leading to tissue damage or organ dysfunction. Some examples are
1. Hashimoto's Thyroiditis: An autoimmune disorder affecting the thyroid gland,
resulting in hypothyroidism. The immune system produces antibodies targeting
thyroglobulin and thyroid peroxidase, impairing hormone synthesis.
2. Autoimmune Anemia: Includes conditions like autoimmune hemolytic anemia,
where antibodies attack red blood cells. Leads to anemia with symptoms like
fatigue, pale skin, and shortness of breath.
3. Insulin-Dependent Diabetes Mellitus (Type 1 Diabetes): the immune system
destroys insulin-producing beta cells in the pancreas. Causes hyperglycemia and
reliance on insulin therapy for management.
★ mediated by Stimulating or Blocking Autoantibodies. These conditions arise when
autoantibodies alter the function of receptors or proteins. Some examples are:
1. Graves' Disease: Autoantibodies stimulate the thyroid-stimulating hormone
(TSH) receptor, leading to excessive thyroid hormone production
(hyperthyroidism).
2. Myasthenia Gravis: Autoantibodies block the acetylcholine receptor at the
neuromuscular junction, impairing communication between nerves and muscles.
Characterized by muscle weakness, particularly affecting the eyes, face, and
swallowing.
B. Systemic autoimmunity
It refers to autoimmune conditions in which the immune system targets ubiquitous or
widely distributed self-antigens. Unlike organ-specific autoimmunity, which is confined to a
single tissue, systemic autoimmune diseases affect multiple organs and tissues simultaneously or
sequentially. Pathogenesis of Systemic Autoimmunity involves:

★ Central Tolerance Failure: Imperfect deletion of self-reactive lymphocytes in the

thymus or bone marrow.
★ Peripheral Tolerance Failure: Dysfunctional regulatory T-cells, impaired anergy,
or faulty apoptosis mechanisms allow self-reactive cells to survive and become
activated.

★ Widespread activation of autoreactive B cells leads to the generation of

autoantibodies, many of which are directed against nuclear components (e.g.,
DNA, histones, RNA-binding proteins).These autoantibodies often form immune
complexes with their target antigens.
★ Circulating antigen–antibody complexes deposit in various tissues (e.g., kidneys,
joints, blood vessels). This leads to activation of the classical complement
pathway, recruitment of inflammatory cells, and local tissue damage —
characteristic of Type III hypersensitivity.

★ As inflammation progresses, damaged tissues release new autoantigens and

aggravates the disease.

★ Overproduction of pro-inflammatory cytokines such as IFN-α, IL-6, TNF-α, and

BAFF (B cell-activating factor) promotes chronic inflammation and perpetuates
autoimmunity.

★ Epigenetic changes (e.g., DNA hypomethylation) can upregulate the expression of

autoantigens or costimulatory molecules.

★ UV radiation, infections, and hormonal influences (e.g., estrogen) can trigger

disease. They may promote apoptosis, increase exposure to nuclear antigens, or
stimulate innate immune pathways (e.g., TLRs recognizing nucleic acids).
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 Some common Autoimmune diseases
Image source: Janeway’s Immunobiology, 9th Edition

IMMUNODEFICIENCY
The immune system is an incredibly adaptable defense mechanism that has evolved to
shield animals from harmful microorganisms and cancer. It can produce a vast array of cells and
molecules that can specifically identify and eliminate an almost endless variety of foreign
threats. These components function together in a highly complex and dynamic network, rivaling
the intricacy of the nervous system.
Immunodeficiency refers to a condition where the immune system is unable to
effectively combat infections. This state is also termed immunocompromised.
Immunodeficiency disorders weaken the immune system’s capacity to protect the body from
foreign or abnormal entities such as bacteria, viruses, fungi, or cancer cells. As a consequence,
individuals may become susceptible to uncommon bacterial, viral, or fungal infections, as well as
lymphomas or other types of cancer. Immunodeficiency reduces the immune system’s ability to
monitor and eliminate cancerous cells, a process known as cancer immunosurveillance.

Primary Immunodeficiency (PID)

Primary immunodeficiency, also referred to as congenital immunodeficiency, is a
condition where an individual's immune system is either underdeveloped or not functioning
properly from birth. Most cases are identified in infants under one year of age. Individuals with
PID are prone to recurring infections, inflammatory conditions, organ dysfunction, and certain
cancers. These disorders are genetic in origin and are categorized into the following types:
✿ B Lymphocyte Deficiency: Disorders arising from B cell deficiencies include conditions
like X-linked agammaglobulinemia (Bruton’s disease) and selective IgA deficiency.

✿ T Lymphocyte Deficiency: T cell-related PIDs include congenital thymic aplasia

(DiGeorge syndrome), chronic mucocutaneous candidiasis, and hyper-IgM syndrome.

✿ Combined B and T Cell Deficiency: Disorders involving both B and T cell impairments
include Severe Combined Immunodeficiency (SCID), Wiskott-Aldrich Syndrome, and
Major Histocompatibility Complex (MHC) deficiency, also known as Bare Lymphocyte
Syndrome.

✿ Phagocyte Deficiency: PIDs due to phagocytic cell deficiencies include chronic

granulomatous disease and leukocyte adhesion deficiency syndrome.

✿ Complement Deficiency: Primary immunodeficiency resulting from complement system

deficiencies can lead to conditions such as hereditary angioedema, frequent infections,
and various autoimmune disorders.

B-Cell Deficiencies include:

(a) X-linked hypogammaglobulinemia
(b) Selective immunoglobulin deficiencies
(c) Hyper-IgM syndrome
(d) Interleukin-12 receptor deficiency

✿ X-linked Hypogammaglobulinemia (XLA): X-linked hypogammaglobulinemia, also

called infantile agammaglobulinemia or XLA, is a classic example of a condition caused
by a lack of B cells. It is usually inherited as a sex-linked disorder, meaning it is passed
 down through the X chromosome. The faulty gene is found at the Xq21.2–22 region,
which controls the production of Bruton’s tyrosine kinase (Btk). Btk is a protein that
helps B cells grow and mature, and it also helps with B-cell signaling later in life.

✿ Selective Immunoglobulin Deficiencies: In this condition, one or more types of

antibodies (immunoglobulins) are missing or low, while the others are normal or even
higher than normal. The most common form is IgA deficiency, where the IgA antibody is
nearly absent in both blood and body secretions. People with IgA deficiency have less
than 5 mg/dL of IgA, but their IgG and IgM levels are usually normal or high. This
condition can run in families or be triggered by infections like measles, other viruses, or
toxoplasmosis. The exact cause of IgA deficiency is unknown, but it is thought to result
from a problem in B cell developmentspecifically, the failure of B cells to become IgA-
producing plasma cells. While IgA-type B cells may exist, they are unable to release IgA.
Since IgA plays a key role in protecting mucosal surfaces (like the lining of the
respiratory and digestive tracts), people with this deficiency often suffer from frequent
respiratory, digestive, and urinary infections. They are also more likely to develop
autoimmune diseases such as lupus and rheumatoid arthritis.
✿ Hyper-IgM Syndrome (HIGM): Hyper-IgM syndrome is a condition where the body
makes too much IgM antibody but very low levels of IgG, IgA, and IgE. People with
this disorder have a normal number of T cells and B cells, but their B cells can’t switch to
making other types of antibodies. Some forms of this condition are inherited through the
X chromosome (X-linked), while others follow an autosomal recessive pattern (inherited
from both parents). People with HIGM are more likely to get repeated infections and may
also develop autoimmune problems like low platelet counts (thrombocytopenia), low
white blood cell counts (neutropenia), and destruction of red blood cells (hemolytic
anemia).

✿ Interleukin-12 Receptor Deficiency: People with this deficiency are very prone to
widespread mycobacterial infections (like tuberculosis). This happens because their
immune cells lack the interleukin-12 (IL-12) receptor, which is needed to trigger a Th1
immune response, a key defense against these types of infections. Without this response,
the body can't fight off mycobacteria effectively.
T- Lymphocyte Deficiency:
Primary immunodeficiency disorders caused by T cell deficiency affect the body’s
ability to fight infections, especially those caused by viruses, fungi, and intracellular bacteria. T
cells play a crucial role in cell-mediated immunity and also help activate B cells to produce
antibodies. When T cells are defective or absent, the immune system becomes severely
compromised. Common T cell-related immunodeficiencies include:
1. Congenital Thymic Aplasia (DiGeorge Syndrome):
Also known as DiGeorge syndrome, this condition results from a developmental defect
in the thymus, the organ where T cells mature. Because the thymus is missing or
underdeveloped, affected individuals have very few or no functional T cells.
Common features include frequent viral and fungal infections due to poor T cell function,
Congenital heart defects, facial abnormalities, Low calcium levels (hypocalcemia) due to
underdeveloped parathyroid glands DiGeorge syndrome is often caused by a deletion on
chromosome 22 (22q11.2 deletion).
2. Chronic Mucocutaneous Candidiasis (CMC):
CMC is a condition marked by persistent and recurrent Candida (yeast) infections,
especially on the skin, nails, and mucous membranes (like the mouth and genitals). It occurs due
to a specific defect in T cell function that impairs the immune response to fungal infections.
Although other parts of the immune system may work normally, the failure of T cells to respond
properly to Candida allows the fungus to grow unchecked.
3. Hyper-IgM Syndrome (linked to T cell signaling defects):
While primarily known for its B cell antibody switching defect, Hyper-IgM syndrome
can also involve problems with T cell function, especially CD40 ligand (CD40L) deficiency on
T helper cells. CD40L is needed to help B cells switch from producing IgM to other antibody
types (like IgG, IgA, or IgE).Without this interaction, IgM levels are abnormally high, other
antibody levels (IgG, IgA, IgE) are very low, Patients have normal T and B cell numbers but
impaired communication between them. This leads to increased susceptibility to bacterial and
fungal infections and a higher risk of autoimmune issues.
Combined B and T Cell Deficiency
Combined immunodeficiencies are a group of primary immunodeficiency disorders
where both B cells (which produce antibodies) and T cells (which coordinate the immune
response and fight infected or abnormal cells) are either missing or not functioning properly.
These are often severe disorders that appear early in life and make affected individuals highly
vulnerable to a wide range of infections. Because both arms of the adaptive immune system are
impaired, patients are at high risk for recurrent bacterial, viral, fungal, and protozoal infections,
Poor response to vaccines, failure to thrive in infancy and increased risk of autoimmune diseases
and cancers. Common examples of combined B and T cell deficiencies include:
1. Severe Combined Immunodeficiency (SCID)
SCID is often called a "bubble boy disease" due to the need for extreme protection from
infections. It is one of the most serious forms of immunodeficiency, appearing in infancy.
Caused by various genetic mutations (X-linked SCID is the most common form), leads to almost
no functional T or B cells, infants appear normal at birth but soon develop severe, recurrent
infections (e.g., pneumonia, chronic diarrhea, thrush) and poor growth and development
2. Wiskott-Aldrich Syndrome (WAS)
WAS is a rare X-linked disorder affecting the immune system and blood cells. Caused
by mutations in the WAS gene, which affects cytoskeletal function in immune cells resulting in
combined immunodeficiency (low T and B cell function), eczema and thrombocytopenia (low
platelet count leading to bleeding)
3. Major Histocompatibility Complex (MHC) Class II Deficiency / Bare Lymphocyte
Syndrome
This rare disorder involves a lack of expression of MHC class II molecules on immune
cells, which is essential for proper T cell activation. Without MHC II, helper T cells (CD4+)
cannot be properly activated, as a result, B cells cannot produce antibodies

Phagocyte Deficiency
Phagocytes are a type of white blood cell that play a key role in the innate immune
system. They detect, engulf (eat), and destroy invading pathogens like bacteria and fungi. The
two main types of phagocytes are neutrophils and macrophages. When phagocytes are
deficient in number or don’t function properly, the body struggles to fight off infections
especially those caused by bacteria and fungi. This condition is known as phagocyte
deficiency, a type of primary immunodeficiency disorder (PID). People with phagocyte
deficiencies often suffer from frequent and severe bacterial infections (especially skin, lungs,
and gastrointestinal tract) delayed wound healing, abscess formation and fungal infections.
Common phagocyte-related immunodeficiencies include:
1. Chronic Granulomatous Disease (CGD): CGD is a genetic disorder where phagocytes
(mainly neutrophils) cannot produce reactive oxygen species (ROS) needed to kill ingested
microbes. As a result, the body tries to isolate infections by forming granulomas (clusters of
immune cells). Patients suffer from repeated skin, lung, liver, and bone infections caused by
bacteria and fungi. Inheritance can be X-linked (most common) or autosomal recessive.
2. Leukocyte Adhesion Deficiency (LAD): In LAD, white blood cells (especially
neutrophils) are unable to move from the bloodstream to the site of infection because they
cannot stick properly to blood vessel walls. This leads to severe bacterial infections, delayed
wound healing, and absence of pus formation despite serious infections.
3. Chediak-Higashi Syndrome: A rare inherited disorder where phagocytes have giant
granules that interfere with their ability to digest pathogens. It is associated with partial
albinism, neurological problems, and recurrent infections. Patients may develop a life-
threatening condition called the accelerated phase, similar to a lymphoma.

Complement Deficiency
The complement system is a group of over 30 proteins in the blood that work together as
part of the innate immune system. These proteins help destroy pathogens, enhance
inflammation, and assist antibodies and phagocytes in clearing microbes from the body. When
there is a deficiency or dysfunction in one or more complement proteins, the immune system
cannot respond properly to infections, especially those caused by bacteria. This condition is
known as complement deficiency, and it is a form of primary immunodeficiency disorder
(PID). Depending on which component is missing, the effects can vary. Complement
deficiencies are generally classified based on which part of the pathway is affected into the
classical pathway, alternative pathway, lectin pathway, or terminal pathway.
Secondary immunodeficiencies
They arise as a result of various diseases, medical conditions, or treatments that suppress
or impair the immune system. Unlike primary immunodeficiencies, which are inherited,
secondary forms are acquired and are far more common. They typically develop due to
underlying pathological processes that compromise immune function or from the use of
medications with immunosuppressive properties. Secondary immunodeficiencies may be
categorized as (a) B-cell deficiencies, (b) T-cell deficiencies, (c) complement deficiencies, and
(d) phagocytic deficiencies. The most prevalent and well-known example is Acquired
Immunodeficiency Syndrome (AIDS), which is caused by infection with the Human
Immunodeficiency Virus (HIV) and leads to a progressive decline in immune competence. In
addition to AIDS, secondary immunodeficiency can result from:
 Immunosuppressive therapies, such as chemotherapy, corticosteroids, or NSAIDs
 Psychological stress and depression
 Severe burns
 Radiation exposure
 Neurodegenerative conditions like Alzheimer’s disease
 Autoimmune and inflammatory diseases, including celiac disease and sarcoidosis
 Hematologic disorders like lymphoproliferative disease, Waldenström’s
macroglobulinemia, multiple myeloma, aplastic anemia, and sickle cell disease
 Nutritional deficiencies
 Aging
 Malignancies
 Chronic illnesses such as diabetes mellitus
 General introduction to vaccines, Types of
vaccines, Immunization programme
Vaccines are one of humanity’s most powerful tools against infectious diseases,
transforming global health and saving countless lives over the centuries. They have shaped the
way societies combat illness, reduce mortality, and improve public health. This essay provides a
detailed overview of vaccines, their types, mechanisms, significance, and future potential. At
their core, vaccines are biological preparations designed to provide immunity against specific
infectious diseases. They achieve this by training the immune system to recognize and combat
pathogens, such as bacteria and viruses. Unlike treatments that address symptoms after an
infection occurs, vaccines work proactively to prevent the disease from taking hold in the body.
The principle of vaccination is the stimulation of an immune response. Vaccines typically
introduce antigens harmless fragments or weakened forms of the pathogen into the body. This
stimulates the immune system to produce antibodies and memory cells, equipping it to recognize
and destroy the actual pathogen if it is encountered in the future. The process of immunization
prepares the immune system by exposing it to a simulated threat, the body is prepared to mount a
rapid and effective defense when faced with the real one. This preventive mechanism forms the
basis of individual immunity and contributes to the broader concept of herd immunity. Herd
immunity, achieved when a large proportion of a population is vaccinated, provides indirect
protection to those who cannot receive vaccines due to medical reasons.
Vaccines undergo rigorous testing in clinical trials before they are approved for public
use. These trials assess their safety and effectiveness, and post-approval surveillance ensures that
any rare adverse effects are monitored and addressed. Mild side effects, such as fever or soreness
at the injection site, are common and signify the immune system’s active response.

History of Vaccines
The history of vaccines is a testament to scientific ingenuity and dedication to public
health. The journey began with variolation in ancient China during the 10th century, where
powdered smallpox scabs were used to prevent the disease. This practice spread to Asia, Africa,
and later Europe, with Lady Mary Wortley Montagu introducing it to England in the 18th
century. The modern era of vaccination emerged in 1796, when Edward Jenner discovered that
cowpox exposure provided immunity against smallpox, coining the term "vaccine." Jenner's
work laid the foundation for immunology. The 19th century saw major advances, including
Louis Pasteur's development of rabies and anthrax vaccines and breakthroughs in bacteriology
that led to vaccines for cholera and typhoid fever.
The 20th century ushered in the "Golden Age of Vaccines," marked by the creation of
polio vaccines by Jonas Salk and Albert Sabin, the eradication of smallpox in 1980, and the
introduction of vaccines for diseases like measles, mumps, rubella, and hepatitis B. In the 21st
century, innovations like mRNA technology brought rapid solutions, as seen with COVID-19
vaccines. Global health initiatives continue to focus on eradicating diseases like polio and
combating outbreaks of Ebola and Zika, showcasing the ongoing impact of vaccines.

Principle of Vaccination
The principle of vaccination is rooted in harnessing the body's natural immune system to
provide protection against specific diseases. It is based on the concept of stimulating an immune
response without causing illness, thereby equipping the immune system to fight off the real
pathogen when encountered. Some key principles include:
1. Stimulating the Immune System
Vaccines introduce antigens either weakened, killed, or harmless fragments of
pathogensinto the body. These antigens act as foreign invaders, prompting the immune system to
recognize the pathogen as a threat. They generate targeted immune response, including the
production of antibodies and activation of immune cells. The antigens presented in a vaccine are
designed to mimic those found on the surface of the actual pathogen, ensuring that the immune
response will be effective if the pathogen is encountered.
2. Formation of Immunological Memory
One of the fundamental principles of vaccination is the creation of immunological
memory. During the immune response, specialized cells called memory B cells and memory T
cells are formed. These memory cells persist in the body for extended periods sometimes for a
lifetime and "remember" the specific antigens. If the individual is exposed to the actual pathogen
later, these memory cells enable the immune system to respond rapidly and efficiently, often
preventing the onset of illness. Immunological memory makes vaccines a powerful tool for long-
term disease prevention.
3. Types of Immune Responses
Vaccines can trigger various types of immune responses. Humoral Immunity involves
the production of antibodies by B cells. Antibodies neutralize pathogens or mark them for
destruction by other immune cells. Cell-Mediated Immunity involves T cells, which destroy
infected cells and regulate other immune responses. This is particularly important for
intracellular pathogens, like viruses. Effective vaccines often stimulate both types of immunity
for comprehensive protection.
4. Herd Immunity
Another key principle of vaccination is herd immunity, which protects populations by
reducing the spread of a pathogen. When a significant portion of a community is vaccinated, the
pathogen's ability to spread is curtailed. This protects individuals who cannot be vaccinated, such
as newborns or those with weakened immune systems. Herd immunity underscores the collective
responsibility of vaccination, highlighting its impact beyond the individual.
5. Safety and Controlled Exposure
Vaccines work by providing controlled exposure to antigens, avoiding the risks
associated with actual infection. This controlled exposure ensures that the immune response is
strong enough to build immunity. The risk of adverse effects is minimized through careful
vaccine design and rigorous testing.
6. Boosting Immunity Through Multiple Doses
For some vaccines, achieving full immunity requires multiple doses or booster shots.
Primary Series involves initial doses that help establish the immune response and create
memory cells. Boosters reinforce immunity by re-exposing the immune system to the antigen.
Boosters are particularly important for vaccines that provide immunity against pathogens with
rapidly mutating antigens or waning immune responses over time.

Types of Vaccines
Vaccines can be categorized based on their formulation into various types, including
whole organism vaccines (either attenuated or inactivated), vaccines derived from purified
macromolecules, recombinant vaccines, DNA-based vaccines, and multivalent subunit vaccines.
1. Live Attenuated Vaccines
Live attenuated vaccines utilize pathogens that have been weakened, so they cannot cause
disease in healthy individuals. These pathogens closely resemble the natural infection, allowing
the immune system to respond robustly and effectively. The attenuated pathogen replicates
minimally within the body, creating an immune response similar to that induced by natural
infection. This stimulates both humoral immunity (antibody production by B cells) and cell-
mediated immunity (activation of T cells).
Examples: Measles, Mumps, Rubella (MMR), Varicella Vaccine, Yellow Fever Vaccine
Advantages:
★ Produces a strong and long-lasting immune response due to its similarity to natural
infections.
★ Often requires fewer doses or boosters for effective immunity.
★ Provides broad immunity, sometimes offering protection against related strains.
Disadvantages:
 Not suitable for people with weakened immune systems, as their bodies might struggle to
control even the weakened pathogen.
 Requires strict storage conditions, such as refrigeration, which can limit accessibility in
resource-poor settings.
 Rarely, the attenuated pathogen can revert to a more virulent form, although this is
exceedingly uncommon with modern vaccines.
2. Inactivated Vaccines
Inactivated vaccines consist of pathogens that have been killed or rendered inactive using
methods like heat or chemicals. These vaccines are incapable of causing disease. Since the
pathogen cannot replicate, the immune response is stimulated by its structural components, such
as proteins or carbohydrates.
Examples: Polio Vaccine (IPV), Hepatitis A Vaccine, Rabies Vaccine
Advantages:
★ Extremely safe for all individuals, including those with compromised immune systems.
★ Stable under normal storage conditions, making them easier to transport and distribute.
Disadvantages:
 Induces a weaker immune response compared to live attenuated vaccines, often requiring
multiple doses or boosters to achieve long-lasting immunity.
 Focuses mainly on humoral immunity, with limited activation of cell-mediated
immunity.
3. Subunit, Recombinant, and Conjugate Vaccines
These vaccines target specific parts of the pathogen, such as proteins, polysaccharides, or
surface antigens. They provide focused immunity without introducing the whole organism. By
isolating critical antigens, these vaccines stimulate the immune system to produce targeted
antibodies against the pathogen.
Examples: Hepatitis B Vaccine, Human Papillomavirus (HPV) Vaccine, Pneumococcal
Conjugate Vaccine
Advantages:
★ Lower risk of side effects because they exclude the pathogen’s unnecessary components.
★ Safe for use in immunocompromised individuals.
★ Highly effective for preventing diseases caused by bacterial toxins or surface antigens.
Disadvantages:
 Generally requires multiple doses or booster shots for sustained immunity.
 Limited in scope, as they only target specific parts of the pathogen, which may not
provide protection against all its strains.
4. Toxoid Vaccines
Toxoid vaccines specifically target diseases caused by bacterial toxins rather than the
bacteria themselves. These vaccines are developed by inactivating the toxins (toxoids) through
chemical or physical means, rendering them harmless while maintaining their ability to stimulate
an immune response. When certain bacteria cause disease, it is often due to the toxins they
release rather than the bacteria themselves. Toxoid vaccines expose the immune system to these
inactivated toxins, teaching it to neutralize them in the event of future exposure.
Examples: Tetanus Vaccine, Diphtheria Vaccine
Advantages
★ Highly effective against toxin-mediated diseases.

★ Safe for most populations, including immunocompromised individuals.

★ Stable formulations make them easy to store and transport.

Disadvantages
 Immunity may weaken over time, requiring booster shots to maintain long-term

protection.
 Limited to combating diseases caused by toxins, not the bacteria themselves.
5. mRNA Vaccines
mRNA vaccines represent a groundbreaking advancement in immunology, using
synthetic messenger RNA to guide the body’s cells in producing a harmless antigen, typically a
protein from the pathogen. This antigen prompts the immune system to develop a defense
mechanism. These vaccines deliver mRNA into the body, which is absorbed by immune cells.
The cells use this mRNA as a template to produce a piece of the pathogen (e.g., the spike protein
of SARS-CoV-2). This triggers an immune response without exposing the individual to the
actual pathogen.
Examples: Pfizer-BioNTech COVID-19 Vaccine, Moderna COVID-19 Vaccine
Advantages
★ Highly efficient at inducing strong immune responses, including both humoral and cell-

mediated immunity.
★ Can be quickly adapted for emerging diseases or variants.

★ Do not use live pathogens, making them extremely safe.

Disadvantages
 Require ultra-cold storage to remain stable, posing challenges for distribution in
resource-limited areas.
 Being a newer technology, there is limited long-term data on their duration of immunity
and potential effects.
6. Viral Vector Vaccines
These vaccines rely on a harmless carrier virus (vector) to deliver genetic material from
the target pathogen into the body. The host cells then produce the antigen, leading to an immune
response. The vector virus is engineered to contain genetic instructions for creating a key protein
from the pathogen. Once inside the body, the vector infects cells, causing them to produce this
protein and trigger immunity.
Examples: AstraZeneca COVID-19 Vaccine, Ebola Vaccine
Advantages
★ Mimic natural infections, eliciting a strong immune response.

★ Effective across various pathogens, making them versatile.

★ Stable under less stringent storage conditions compared to mRNA vaccines.

Disadvantages
 Pre-existing immunity to the vector virus in some individuals may reduce the
effectiveness of the vaccine.
 Rare but serious adverse events, such as blood clotting disorders, have been reported.
7. DNA Vaccines
DNA vaccines use small circular DNA molecules (plasmids) that encode antigens. These
plasmids instruct the host cells to produce the antigen, triggering an immune response. The DNA
plasmid is introduced into cells, usually via injection. The host cells use this genetic material to
produce the pathogen's antigen, which is then presented to the immune system to elicit
immunity.
Advantages
★ Simple to manufacture and relatively low-cost.

★ Stable at room temperature, simplifying storage and distribution.

★ Can be quickly developed for emerging diseases.

Disadvantages
 Limited usage and clinical approval compared to other vaccine types.
 May elicit a weaker immune response, necessitating booster doses or other
enhancements.
8. Combination Vaccines
Combination vaccines merge antigens from multiple pathogens into one formulation,
reducing the number of injections required. By integrating multiple antigens in a single shot,
these vaccines simplify immunization schedules and enhance vaccination compliance, especially
in children.
Examples : DTaP Vaccine -Protects against diphtheria, tetanus, and pertussis.
Advantages
★ Reduces the burden of multiple injections, particularly for infants and children.

★ Increases compliance with immunization programs.

★ Convenient for both individuals and healthcare systems.

Disadvantages
 The combination of antigens may lead to stronger immune reactions in some individuals.
 Complex formulations can increase manufacturing challenges.
Emerging and Experimental Vaccine Technologies

⬥ Nanoparticle Vaccines: Use nanoparticles as delivery systems to efficiently present

antigens to the immune system.

⬥ Personalized Cancer Vaccines: Target unique mutations in an individual’s tumor,

offering personalized treatment options.

⬥ Universal Vaccines: Aim to provide broad immunity against highly variable

pathogens like influenza.

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National Immunisation Schedule
 Universal Immunisation schedule in India

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 Organ transplantation – graft rejection and
immunosuppressors

Transplantation refers to the medical procedure in which cells, tissues, or organs are
surgically relocated from one part of the body to another, either within a single individual or
between a donor and a recipient. This method is a critical intervention employed when an organ
or tissue is no longer able to function adequately due to damage caused by disease, injury, or
failure. Through transplantation, a damaged organ system can be replaced with a healthy
counterpart from a donor, restoring essential functions and improving the recipient's quality of
life.
Organ transplantation is a complex and major surgical operation, generally considered a
last resort when all other medical treatments have proven insufficient. Due to its intricacies, it
needs meticulous planning and execution, as well as careful post-operative monitoring to ensure
long-term success.
The immune system plays an indispensable yet challenging role in the transplantation
process. While its primary function is to detect and eliminate harmful foreign invaders such as
bacteria and viruses, these same immune mechanisms can unintentionally become a significant
obstacle. The transplanted organ or tissue, referred to as the graft, is often recognized as a
foreign entity by the recipient's immune system. This triggers a defense mechanism known as
transplant rejection, in which the immune system attacks the graft, ultimately leading to its
destruction if left unmanaged.
The severity of the immune response largely depends on factors such as the type of graft
being used and the genetic differences between the donor and recipient. For example, grafts with
greater genetic disparities are more likely to provoke strong immune reactions, increasing the
risk of rejection. To mitigate this risk, donor and recipient matching is conducted prior to
transplantation. By carefully assessing immune compatibility, healthcare professionals aim to
minimize genetic discrepancies and reduce the likelihood of rejection.
Advancements in immunosuppressive medications further enhance the success rate of
transplantations by inhibiting the immune response and allowing the graft to integrate with the
recipient's body. But, these treatments necessitate a delicate balance to avoid complications, such
as increased vulnerability to infections. Transplantation represents a remarkable achievement in
modern medicine, offering patients with end-stage organ failure or irreversible tissue damage the
possibility of renewal and recovery. The surgical expertise and immunological considerations
emphasizes the complexity and significance of this lifesaving procedure.

Transplants or Grafts are categorized based on the source of the transplanted tissue or organ:

1. Allografts: These involve the transfer of tissue from one individual to another within the
same species. The individuals are genetically distinct, which may lead to immune
responses or rejection if precautions are not taken. Allografts are commonly used in
procedures like kidney transplants.
2. Isografts: It a specialized type of allograft which include transplanted tissues between
identical twins. Since monozygotic twins are genetically identical, the likelihood of
rejection is minimal, making these transplants highly successful.
3. Autografts: These are transplants where tissue is moved from one location on an
individual's body to another. For instance, skin grafts applied to burn wounds or the use
of tissue from one part of the body to reconstruct another part fall under this category.
Autografts eliminate the risk of immune rejection as the tissue comes from the same
individual.
4. Xenografts: This category involves the transplantation of tissue or organs from one
species to another, such as transferring tissues from animals to humans. While xenografts
have unique challenges, including immunological barriers, they are being explored in
medical research and applications like heart valve replacements derived from pigs.
 Types of grafts
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Immunology of Transplant Rejection

Transplant rejection is a complex immunological process. It is influenced by the ability of
the immune system to distinguish between what belongs to the body (self) and what is foreign
(non-self) to the body. This feature is fundamental to maintain health, prevent infections, and
avoid autoimmune diseases where the immune system mistakenly attacks its own tissues.
Distinguishing Between Self and Non-Self
The immune system employs highly specialized mechanisms to recognize and eliminate
foreign invaders like bacteria, viruses, or foreign cells. These invaders are identified through
specific molecules called antigens, which act as markers on the surface of pathogens or foreign
cells. Upon detecting non-self antigens, the immune system activates its defense mechanisms,
which include producing antigen-specific antibodies that bind to infected cells, marking them for
destruction while amplifying the immune response. At the core of self-recognition lies the
Human Leukocyte Antigen (HLA) complex, a set of genes responsible for encoding proteins
that serve as "identity markers" for the immune system. These proteins are displayed on the
surface of all cells in the body, signaling to the immune system that these cells are part of the
individual's self, and thus should not be targeted for destruction. Each individual possesses a
unique set of HLA markers based on their genetic makeup, which the immune system learns to
recognize and tolerate. However, any cell or tissue lacking the individual's specific HLA markers
will be considered as foreign and subjected to an immune attack.
Mechanism of Transplant Rejection:
Graft rejection occurs when the immune system of the recipient identifies the
transplanted organ or tissue (graft) as foreign and initiates an immune response against it. This
response is primarily triggered by the recognition of the donor's HLA proteins, which differ from
those of the recipient. These differences lead the recipient's immune cells to perceive the graft as
a threat and mobilize a defense, ultimately compromising the viability of the transplanted tissue.
A critical factor in determining the intensity of this immune response is histocompatibility,
which refers to the genetic compatibility between the donor and recipient based on their HLA
profiles. Greater compatibility results in reduced immune reactions, whereas significant genetic
disparity increases the likelihood of rejection. Even in cases of high compatibility, such as
identical twins (isografts), rejection is still possible due to other donor-specific antigens present
on the graft.
Another complexity arises with certain grafts, such as bone marrow transplants, where
mature immune cells present in the donor graft can recognize the recipient's cells as foreign. This
phenomenon, known as graft-versus-host disease (GVHD), involves the donor's immune-
competent cells attacking the recipient's tissues. GVHD is a significant risk in stem cell
transplants and can also occur after blood transfusions containing immune-reactive cells.
Important components involved in graft rejection:
The process of antigen presentation is the critical initial step in graft rejection. The
proteins encoded by the donor's Human Leukocyte Antigen (HLA) complex, along with other
donor-specific antigens, are recognized by the recipient's immune system as foreign. Specialized
immune cells, known as antigen-presenting cells (APCs)—such as dendritic cells,
macrophages, or B cells—play a pivotal role here. These APCs process the foreign donor
antigens and present them on their surfaces using Major Histocompatibility Complex (MHC)
molecules. The MHC-peptide complexes are then displayed to the recipient's T cells in the
lymphoid organs. This step is essential for initiating the adaptive immune response, and it
determines the strength and nature of the recipient’s reaction to the graft.
A) T Cell Activation
Once the donor antigens are presented, the recipient’s T cells—particularly cytotoxic T
cells (CD8+ T cells) and helper T cells (CD4+ T cells)—are activated. Cytotoxic T cells are
directly involved in attacking the graft by inducing apoptosis (programmed cell death) in donor
cells. Meanwhile, helper T cells release pro-inflammatory cytokines, such as interleukin-2 (IL-
2) and interferon-gamma (IFN-γ), which amplify the immune response by recruiting additional
immune cells to the site of the graft.
This activation occurs in a sequence of events such as
1. Recognition: The T cell receptor (TCR) on the recipient's T cells specifically binds to the
MHC-antigen complex presented by APCs.
2. Costimulation: Additional signals provided by costimulatory molecules (e.g., CD28 on
T cells binding to B7 on APCs) ensure full T cell activation.
3. Proliferation and Differentiation: The activated T cells proliferate and differentiate into
effector T cells that mediate the immune response.

B) B Cell Involvement
B cells are another key player in graft rejection, particularly in antibody-mediated
rejection (AMR). These cells become activated after recognizing donor antigens and interacting
with helper T cells. Once activated, B cells differentiate into plasma cells, which produce
antibodies specific to the donor antigens. These antibodies can bind to donor cells, target them
for destruction via mechanisms such as complement activation and also recruit additional
immune cells, such as natural killer (NK) cells, through a process known as antibody-
dependent cellular cytotoxicity (ADCC). The production of these donor-specific antibodies
(DSAs) can lead to vascular inflammation and damage, playing a central role in acute and
chronic graft rejection.

Effector Mechanisms
Effector mechanisms include the various ways in which immune cells and antibodies
mediate the destruction of the graft. These mechanisms include:
1. Direct Cell Lysis: Cytotoxic T cells induce apoptosis in donor cells by releasing
cytotoxic molecules such as perforin (which creates pores in the target cell membrane)
and granzymes (which trigger cell death).
 2. Inflammation: Helper T cells and other immune cells release inflammatory cytokines
(e.g., TNF-α, IL-1), which recruit macrophages and neutrophils to the graft site. These
cells exacerbate tissue injury by producing reactive oxygen species (ROS) and proteolytic
enzymes.
3. Antibody-Mediated Damage: Antibodies bind to endothelial cells lining the graft’s
blood vessels, triggering the activation of the complement cascade. This process results in
the formation of the membrane attack complex (MAC), which disrupts cell membranes
and damages graft vasculature. The complement system also generates inflammatory
mediators (e.g., C3a, C5a) that further amplify the immune response.
4. Vascular Disruption: The immune response against the graft's blood vessels can lead to
thrombosis (clot formation) and loss of blood supply, causing ischemia and ultimately
graft failure.

Immunological basis of graft rejection

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Stages of Transplant Rejection

Transplant rejection is categorized into distinct stages based on the timing, mechanism,
and severity of the immune response. Each stage presents unique challenges that require specific
interventions to protect the viability of the graft.
Hyperacute Rejection
It is the most rapid form of transplant rejection, occurring within minutes to hours after
the transplantation. This swift immune response is triggered by preexisting antibodies in the
recipient that specifically recognize foreign antigens present on the donor's graft. These
antibodies may have been generated due to previous blood transfusions, earlier transplants, or
multiple pregnancies, where the immune system encountered and "memorized" foreign antigens.
When the graft is transplanted, these antibodies bind to the antigens on the blood vessels
of the graft, leading to the formation of blood clots. These clots obstruct blood flow, preventing
oxygen and nutrients from reaching the graft. As a result, the graft is severely compromised and
rejected almost immediately. Hyperacute rejection is irreversible and requires the prompt
removal of the graft to prevent further complications.
Acute Rejection:
It is a common stage that typically occurs within the first 6 months following
transplantation. While the risk is highest during the first 3 months, rejection can arise at later
intervals as well. This stage is caused by the recipient's immune system detecting non-self
antigens in the donor graft, prompting the formation of antibodies and the activation of immune
cells against the graft.
Unlike hyperacute rejection, acute rejection often develops gradually and can be
effectively managed if identified early. Immunosuppressive treatments are used to inhibit the
immune response, reducing inflammation and damage to the graft. In many cases, these
interventions can preserve the function of the transplanted organ or tissue and prevent
irreversible damage.
Chronic Rejection
It represents the long-term failure of the graft and generally occurs months to years after
transplantation. This stage is often the result of repeated episodes of acute rejection, which
gradually impair the structure and function of the graft. Chronic rejection is characterized by
scarring and fibrosis, processes that progressively deteriorate the transplanted tissue or organ.
Unlike hyperacute and acute rejection, chronic rejection is a slow and dangerous process
that is difficult to treat effectively. There are currently no curative treatments for chronic
rejection, and the only option in advanced cases may be the removal and replacement of the
graft. Managing chronic rejection focuses on minimizing inflammation and slowing the
progression of damage through immunosuppressive therapies.

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Immunosuppressors in Graft Rejection
Immunosuppressors are essential in transplantation medicine to prevent or manage graft
rejection. These medications work by suppressing the recipient's immune system, reducing its
ability to attack the transplanted organ or tissue.
Phases of Immunosuppression
1. Induction Therapy: This phase aims to prevent early rejection immediately after the
transplant procedure when the risk of an acute immune response is highest. Induction
therapy involves the use of high-intensity immunosuppressive agents, such as
monoclonal antibodies (e.g., Basiliximab) or polyclonal antibodies (e.g., Antithymocyte
Globulin). These agents act to deplete or inhibit T cells, which are central to the rejection
process. This therapy allows the graft to begin integration with the recipient’s body while
minimizing early immune attacks.
2. Maintenance Therapy: Following the critical post-transplantation period, this long-term
therapy is used to sustain graft function and prevent both acute and chronic rejection.
Lower doses of immunosuppressive drugs are administered to minimize side effects. The
goal is to balance immune suppression with minimizing risks like infections or other
complications arising from prolonged use of immunosuppressors. This therapy typically
involves a combination of:
★ Calcineurin inhibitors (e.g., Cyclosporine, Tacrolimus).

★ Antiproliferative agents (e.g., Mycophenolate Mofetil, Azathioprine).

★ Corticosteroids (e.g., Prednisone).

3. Treatment of Rejection: When signs of acute or chronic rejection are detected, more
aggressive strategies are employed to counteract the immune response and restore graft
function. High doses of corticosteroids, monoclonal or polyclonal antibodies, and
adjustments to maintenance therapy regimens are used during this phase. The treatments
aim to target activated immune cells and suppress inflammation.

Important Immunosuppressors:
Immunosuppressive drugs are essential in managing graft rejection, as they inhibit or
modulate the recipient's immune system to ensure the survival and functionality of the
transplanted organ or tissue. Each class of immunosuppressors targets specific pathways in the
immune system, and they are carefully combined to achieve optimal therapeutic outcomes.
1 Calcineurin Inhibitors: Calcineurin inhibitors block the activity of the enzyme
calcineurin, which is crucial for activating T cells. Calcineurin normally facilitates the
production of interleukin-2 (IL-2), a cytokine that promotes T cell proliferation. By
inhibiting calcineurin, these drugs effectively suppress the expansion of activated T cells,
a key component in the immune response against the graft. These are foundational drugs
in maintenance therapy and are highly effective in preventing acute rejection by limiting
T cell-mediated immune responses. Long-term use can lead to nephrotoxicity (kidney
damage), hypertension, increased cholesterol levels, and a heightened risk of infections.
Patients require frequent monitoring of blood levels to optimize dosing and minimize
toxicity.
Examples: Cyclosporine, Tacrolimus.
2 Corticosteroids: Corticosteroids reduce inflammation by inhibiting the production of
pro-inflammatory cytokines such as TNF-α, IL-1, and IL-6. They also interfere with the
activation and functioning of immune cells, including T cells, B cells, and macrophages,
thereby dampening the immune response against the graft. Corticosteroids are versatile
and used in all phases of immunosuppression - induction therapy, maintenance therapy,
and treatment of acute rejection episodes. High doses are especially effective in reversing
acute rejection. Long-term use can result in hyperglycemia (high blood sugar),
osteoporosis (weakened bones), weight gain, cataracts, and increased susceptibility to
infections.
Examples: Prednisone, Methylprednisolone.
3 Antiproliferative Agents: These drugs inhibit the synthesis of nucleotides necessary for
lymphocyte (T and B cell) proliferation. Mycophenolate Mofetil (MMF) targets inosine
monophosphate dehydrogenase, a key enzyme in purine synthesis, selectively inhibiting
lymphocyte proliferation. Azathioprine acts as a purine analog, interfering with DNA
and RNA synthesis, broadly suppressing the proliferation of immune cells. Commonly
used as part of maintenance therapy alongside calcineurin inhibitors to provide additional
suppression of immune responses. Side effects include bone marrow suppression, leading
to reduced blood cell counts, gastrointestinal disturbances (nausea, diarrhea), and a higher
risk of infections.
Examples: Mycophenolate Mofetil (MMF), Azathioprine.
4 mTOR Inhibitors: mTOR inhibitors block the mammalian target of rapamycin (mTOR),
a protein kinase involved in cell growth, proliferation, and metabolism. By inhibiting
 mTOR, these drugs suppress the activation and proliferation of T cells and other immune
cells. It is an alternative to calcineurin inhibitors, particularly useful for patients
experiencing nephrotoxicity from traditional regimens. They also promote anti-fibrotic
effects, reducing scarring in the transplanted organ. Delayed wound healing,
hyperlipidemia (high cholesterol levels), and increased susceptibility to infections are
common side effects. mTOR inhibitors require careful monitoring in post-operative
settings.
Examples: Sirolimus (Rapamycin), Everolimus.
5 Monoclonal Antibodies: Monoclonal antibodies are engineered to target specific
molecules on immune cells. Basiliximab and Daclizumab block the IL-2 receptor on T
cells, preventing their activation and proliferation. Rituximab targets CD20 on B cells,
depleting B cells and reducing antibody-mediated rejection. These antibodies are widely
used in induction therapy and the treatment of acute rejection cases, particularly in
antibody-mediated rejection (AMR). Side effects include infusion-related reactions,
increased infection risk, and potential allergic responses during administration.
Examples: Basiliximab, Daclizumab, Rituximab.
6 Polyclonal Antibodies: Polyclonal antibodies target multiple antigens on T cells and
other immune cells, leading to their depletion and suppression of immune responses.
ATG specifically depletes lymphocytes, while Alemtuzumab targets CD52, a surface
marker on lymphocytes. It is highly effective for induction therapy and treating severe
rejection cases. They are particularly useful in patients at high risk for rejection.
Associated with cytokine release syndrome, bone marrow suppression, and increased
infection risk.
Examples: Antithymocyte Globulin (ATG), Alemtuzumab.
7 Costimulation Blockers: Belatacept blocks the interaction between CD28 on T cells and
B7 on antigen-presenting cells (APCs), preventing the activation of T cells. This
innovative mechanism bypasses traditional pathways, reducing dependency on
calcineurin inhibitors. It is an alternative for maintenance therapy, particularly in patients
who cannot tolerate calcineurin inhibitors. Increased risk of infections and post-transplant
lymphoproliferative disorder (PTLD) are significant concerns.
Examples: Belatacept.
 About the Author
Dr. Y. Shanti Prabha
Senior Lecturer, Department of Zoology
Dr. V.S. Krishna Government Degree and PG College (Autonomous), Visakhapatnam, India

Academic Background and Specialization

Dr. Y. Shanti Prabha is a distinguished academic in the field of Zoology, with a
specialized focus on Fisheries Science. She completed her postgraduate studies at Andhra
University, securing the first rank and earning a gold medal for her exceptional performance. She
later obtained her Ph.D. from the same university, where she conducted pioneering research in
fisheries-related areas. Her academic excellence has been recognized through several awards,
including the prestigious Nyapati Lakshmi Srinivas Iyyengar Prize and the Jayanthi Venkubai
Memorial Medal.
Research Contributions and Publications
Dr. Shanti Prabha maintains an active research profile with numerous publications in
reputed national and international journals. Her research interests include fish nutrition, bioactive
peptides derived from marine organisms, and chemical quality indices in seafood. She has
successfully guided multiple student research projects and currently supervises postgraduate and
doctoral candidates.
Teaching and Academic Involvement
With over a decade of teaching experience, Dr. Shanti Prabha brings expertise in
Zoology, Environmental Science, Fisheries and Aquaculture. She employs a student-centered
teaching methodology that integrates technology, interactive learning, and practical lab sessions.
She has been instrumental in designing undergraduate curricula and serves on various Boards of
Studies, helping to shape academic programs across institutions.
Professional Development and Authorship
Dedicated to professional growth, she regularly attends workshops, faculty development
programs, and international conferences. She has also authored practical manuals and textbook
chapters for the School of Distance Education at Andhra University, covering a broad spectrum
of topics within Zoology and its allied disciplines. Dr. Y. Shanti Prabha is recognized for her
academic excellence, research contributions, and unwavering commitment to education and
community service.