Chapter 3: Fundamentals of
Immunology
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Introduction
Cellular defense mechanism
• mediated by macrophages, T cells, and dendritic
cells.
Humoral mechanism
• B cells produce specific antibodies
• Complement binds to immunoglobulin molecules
that have specific complement receptor sites.
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Introduction (cont’d)
Detection of allo- and auto- antibodies or
• depends upon binding forces between antigens
and antibodies, properties of the antibody itself,
and individual host characteristics.
• Antigen-antibody reactions are influenced by a
number of factors, including distance, antigen-
antibody ratio, pH, temperature, and
immunoglobulin type
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Overview of the Immune System
Innate or natural immunity
Acquired or adaptive immunity
Cellular and humoral immunity
• Cellular defense mechanism: cells involved
• Humoral defense mechanisms: antibodies and
complement
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Overview of the Immune System (cont’d)
Types of Antibodies
• Autoantibodies: directed against self antigens
• Alloantibodies: directed vs. non-self antigens
• Antigen: molecule found on the surface of
foreign cells or on damaged internal cells
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Overview of the Immune System: Cells
B cells
T cells
Antigen-presenting cells
NK cells
Immune system organs
• Primary lymphoid organs
• Secondary lymphoid organs
MHC Class I and Class II antigens
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Overview of the Immune System: Cells
(cont’d)
Cell linage markers
• Cluster Designation (CD)
• pluripotent stem cell -- CD 34
• Granulocytes – CD11b, CD16, CD35
• T cells – CD2, CD3, CD4, CD8
• B cells – CD19, CD20, CD21, CD22, CD35
• NK cells – CD16, CD56
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Overview of the Immune System (cont’d)
Importance of cytokines and immunoregulatory
molecules
• lymphokines
• monokines
Basic immune system genetics
Major Histocompatibility Complex (MHC)
• Class I HLA molecules
• Class II HLA molecules
• Role of MHC Class I and Class II molecules
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Characteristics of Immunoglobulins
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Heavy chains
• Gamma (IgG)
<Insert Figure 3-2>
• Alpha (IgA)
• Mu (IgM)
• Delta (IgD)
• Epsilon (IgE)
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Characteristics of Immunoglobulins
(2 of 5)
Light chains
• Kappa
• Lambda
Basic immunoglobulin structure
• Papain and pepsin digestion
• Disulfide bonds
• Constant and variable regions
FC region binds complement
Fab region binds antigen
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Characteristics of Immunoglobulins
(3 of 5)
Immunoglobulins significant for blood banking
• IgG, IgM, and IgA: most significant
• Reaction temperatures
• Naturally occurring antibodies
• Commonly encountered IgM and IgG antibodies
• IgG subclasses
• Role of IgE
• Role of IgD
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Characteristics of Immunoglobulins
(4 of 5)
Immunoglobulins significant for blood
banking
• IgM can exist in monomeric and pentameric
forms
<Insert Figure 3-4>
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Characteristics of Immunoglobulins
(5 of 5)
Immunoglobulin variation
• Isotype
• Allotype
• Idiotype
Immunoglobulin Fc receptors
• IgG subclasses involved
• Cells involved
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Complement System
Biological roles
Mediation of inflammation
Circulating and cell membrane proteins
Activation pathways
• Classical
• Alternative
• Lectin
• Membrane attack complex
• Binding of complement by RBC antibodies
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Complement System (2 of 7)
Nomenclature
• Numbering of components
• Designation of active and fragment components
• Nomenclature unique to the alternative pathway
• Inhibitory and regulatory molecules
Role of calcium and magnesium
Regulation
• Self recognition
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Complement System (3 of 7)
Click icon to add picture
Classical Complement
Pathway
• Activation when antibody
binds to antigen
• Activation of components
<Insert Figures 3-5>
• Fragments with
anaphylatoxin activity
• Membrane attack
complex formation
• Cell lysis
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Complement System (4 of 7)
Alternative complement pathway
Activation of alternative pathway
Important protein factors
• Factor D
• Factor B
• Factor P
• C3
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Complement System (5 of 7)
Lectin Complement Pathway
• Activation
• Elements common with classical pathway
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Complement System (6 of 7)
Membrane Attack Complex (MAC)
• Initiation of MAC in the classical pathway
• Initiation of MAC in the alternative pathway
• Complement mediated cell destruction
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Complement System (7 of 7)
Binding of complement by RBC antibodies
• Activation by IgG
• Activation by IgM
• IgG Rh antibodies
• IgM Lewis antibodies
• ABO antibodies
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Characteristics of Antigens
Initiate formation of and reactions to antibodies
Antigen characteristics influencing immune
response: size, complexity, conformation, charge,
accessibility, solubility, digestibility, chemical
composition
Different blood group antigens differ in their
immunogenicity
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Characteristics of Blood Group Antibodies
Polyclonal antibodies
Monoclonal antibodies
Naturally occurring antibodies
• Produced without transfusion, injection, pregnancy
• IgM, RT or lower, activate complement, may be hemolytic
at 37°C
• ABH, Hh, Ii, Lewis, MN, P
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Characteristics of Blood Group Antibodies
(2 of 4)
Immune antibodies
• Transfusion or pregnancy
• IgG, 37°C
• Require AHG for detection
• Rh, Kell, Duffy, Kidd, Ss
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Characteristics of Blood Group Antibodies
(3 of 4)
Unexpected antibodies
Naturally occurring ABO antibodies
• Isoagglutinins
Unexpected antibodies
• Detection techniques
• Importance in pretransfusion testing
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Characteristics of Blood Group Antibodies
(4 of 4)
Alloantibodies and autoantibodies
Production of alloantibodies
Production of autoantibodies
• Panagglutinins or polyagglutinins
• Effects of temperature
• Presence of autoimmune disease
• Special transfusion techniques needed
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Characteristics of Antigen-Antibody
Reactions
Intermolecular binding <Insert Figure 3-7>
forces
Antibody properties
• Affinity
• Avidity
Antibody specificity
• Specific reaction
• Cross-reaction
• No reaction
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Characteristics of Antigen-Antibody
Reactions (cont’d)
Host factors influencing immune response
Influence of MHC system
Duffy system and malaria
Immune tolerance effects
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Detection of RBC Antigen-Antibody
Reactions
Blood samples required
Different tests may require different samples
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Traditional Laboratory Methods
Hemagglutination
Precipitation
Agglutination Inhibition
Hemolysis
ELISA (EIA), IF, WB
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Red Cell Agglutination Reactions
Sensitization
Lattice formation
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Factors Influencing Agglutination
Reactions
Centrifugation <Insert Figure 3-8>
Zeta potential
Antigen-antibody ratio
• Prozone
• Equivalence
• Postzone
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Factors Influencing Agglutination
Reactions (2 of 3)
Effects of pH
Temperature
Immunoglobulin type
Different techniques for IgG and IgM
Enhancement media – especially for IgG antibodies
Protein media
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Factors Influencing Agglutination
Reactions (3 of 3)
Zeta potential, sialic acid in red cells
Low Ionic Strength Media (LISS)
Polyethylene Glycol (PEG) and polybrene
Proteolytic enzymes
Antihuman Globulin (AHG) reagents
Chemical reduction of IgG and IgM
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Monoclonal versus Polyclonal Reagents
Differences in manufacturing techniques
Disadvantages of monoclonal reagents
• Overspecificity
• Complement may not be fixed in antigen-
antibody reaction
• Oversensitivity
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Nontraditional Laboratory Methods
Flow Cytometry
• Quantify fetomaternal hemorrhage
• Study transfused cells
• Distinguish heterozygous and homozygous
antigen expression
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Diseases Important in Blood Bank
Serologic Testing
Immunodeficiency
Hypersensitivity
Monoclonal and polyclonal gammopathies
Autoimmune disease
Hemolytic Disease of the Newborn (HDN)
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Blood Product Transfusions and the
Immune System
Transfusion-Related Immunomodulation
(TRIM)
• Cells, cytokines involved in TRIM
• Effects of leukoreduction
• Reducing incidence of TRIM
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