Antigens

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 Immunogenicity Versus Antigenicity
• Antigens: Substances that can be recognized by the immunoglobulin receptor of B cells, or by the
T cell receptor are called as antigens, in general.
• Immunogenicity is the ability to induce a humoral and/or cell mediated immune response:

B cells + antigen effector B cells + memory B cells

(plasma cells)

T cells + antigen effector T cells + memory T cells

(e.g., CTLs, THs)

• Though a substance that induces a specific immune response is usually called an antigen, it is
more appropriately called an immunogen.

• Antigenicity is the ability to combine specifically with the final products of the above responses,
e.g. antibodies.

• Although all molecules that have the property of immunogenicity also have the property
of antigenicity, the reverse is not true.

• Some small molecules, called haptens, are antigenic but incapable, by themselves, of inducing a
specific immune response.
• In other words, haptens lack immunogenicity.
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 Factors that influence immunogenicity
Factors that affect immunogenicity are two broad categories

Nature of Biological
immunogen. system.

1. Foreignness.
1. Genotype of recipient
2. Molecular size animal.

3. Chemical composition and 2. Immunogen dosage and

heterogeneity route of administration.

4. Susceptibility to antigen
processing and 3. Adjuvants.
presentation.

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 Nature of antigen
• Foreignness: To start an immune response, a molecule must be recognized as nonself by the
biological system. The capacity to recognize nonself is accompanied by tolerance of self, a
specific unresponsiveness to self antigens.
• When an antigen is introduced into an organism, the degree of its immunogenicity depends
on the degree of its foreignness.

• Generally, the greater the phylogenetic distance between two species, the greater the
structural (and therefore the antigenic) disparity between them.

• Thus, foreignness is directly proportional to the evolutionary distance between

immunogen and its host.

• E.g. 1. Common experimental antigen bovine serum albumin (BSA) is not immunogenic
when injected into a cow but is strongly immunogenic when injected into a rabbit.

2. BSA exhibits greater immunogenicity in a chicken than in a goat, which is more closely
related to bovines.

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 Exceptions to
foreignness rule

Some macromolecules such as collagen, cytochrome c, histones etc. have been highly
conserved during evolution process, thus display very little immunogenicity across diverse
species lines.

Some self components such as sperm and cornea are effectively sequestered from the immune
system, thus they will show strong immunogenicity even in their species of origin.

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• Molecular size: There is a correlation between the size of a macromolecule and its
immunogenicity. The most active immunogens tend to have a molecular mass of 100,000
daltons (Da).

• Generally, substances with a molecular mass less than 5000–10,000 Da are poor
immunogens, although a few substances with a molecular mass less than 1000 Da have
proven to be immunogenic.

• Chemical composition and heterogeneity: Size and foreignness are not, by themselves,
sufficient to make a molecule immunogenic; other properties are needed as well.

For example, synthetic homopolymers (polymers composed of a single amino acid or sugar)
tend to lack immunogenicity regardless of their size.
Studies have shown that copolymers composed of different amino acids or sugars are usually
more immunogenic than homopolymers of their constituents. These studies show that
chemical complexity contributes to immunogenicity.

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 In this regard it is notable that all four levels of protein organization—primary, secondary,
tertiary, and quaternary—contribute to the structural complexity of a protein and hence
affect its immunogenicity.

The development of both humoral and cell mediated immune

Concept responses requires interaction of T cells with antigen that has
been processed and presented together with MHC molecules.

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• Susceptibility to antigen processing and presentation:

• Large, insoluble macromolecules generally are more immunogenic than small,

soluble ones because the larger molecules are more readily phagocytosed and
processed.

Macromolecules that cannot be degraded and presented with MHC molecules are
poor immunogens.

This can be illustrated with polymers of D-amino acids, which are stereoisomers of
the naturally occurring L-amino acids.

Because the degradative enzymes within antigen-presenting cells can degrade only
proteins containing L-amino acids, polymers of D-amino acids cannot be processed
and thus are poor immunogens.

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 The biological system
• Genotype of recipient: The genetic constitution (genotype) of an immunized animal
influences the type of immune response the animal manifests, as well as the degree of the
response.
• Example: Hugh McDevitt experiment that showed that two different inbred strains of mice
responded very differently to a synthetic polypeptide immunogen. After exposure to the
immunogen, one strain produced high levels of serum antibody, whereas the other strain
produced low levels. But why ????

F1 generation showed
intermediate response.

Backcross analysis to map the

genes responsible.

Genes responsible found in

the MHC region.

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• Immunogen dosage and route of administration:
Each experimental immunogen exhibits a particular dose-response curve, which is
determined by measuring the immune response to different doses and different
administration routes.
An antibody response is measured by determining the level of antibody present in the
serum of immunized animals.

• Importance of dosage: An insufficient dose will not stimulate an immune response either
because it fails to activate enough lymphocytes or because, in some cases, certain ranges of
low doses can induce a state of immunologic unresponsiveness, or tolerance.

Conversely, an excessively high dose can also induce tolerance.

Example: The immune response of mice to the purified pneumococcal capsular

polysaccharide illustrates the importance of dose. A 0.5 mg dose of antigen fails to induce an
immune response in mice, whereas a thousand-fold lower dose of the same antigen (0.5μg)
induces a humoral antibody response.

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• Booster dosage: A single dose of most experimental immunogens will not induce a strong
response; rather, repeated administration over a period of weeks is usually required.

• Such repeated administrations, or boosters, increase the clonal proliferation of antigen-

specific T cells or B cells and thus increase the lymphocyte populations specific for the
immunogen.

• Importance of antigen delivery route:

Experimental immunogens are generally administered parenterally (para, around; enteric,
gut)—that is, by routes other than the digestive tract. Routes of administration are:
• Intravenous (iv): into a vein
• Intradermal (id): into the skin
• Subcutaneous (sc): beneath the skin
• Intramuscular (im): into a muscle
• Intraperitoneal (ip): into the peritoneal cavity.

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• Adjuvants: Adjuvants (from Latin adjuvare, to help) are substances that, when
mixed with an antigen and injected with it, enhance the immunogenicity of that
antigen.

• When are they used??? Adjuvants are used to boost the immune response when
1. An antigen has low immunogenicity
2. When only small amounts of an antigen are available.

• How do they function??? Precisely how adjuvants augment the immune response
is not entirely known, but they use one or more of the following effects:

• Antigen persistence is prolonged.

• Co-stimulatory signals are enhanced.
• Local inflammation is increased.
• The nonspecific proliferation of lymphocytes is stimulated.

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• Examples of famous adjuvants:
• Freund’s incomplete adjuvant contains antigen in aqueous solution, mineral oil,
and an emulsifying agent such as mannide monooleate, which disperses the oil
into small droplets surrounding the antigen.
• The antigen is then released very slowly from the site of injection.
• This preparation is based on Freund’s complete adjuvant.

• Freund’s complete adjuvant, the first deliberately formulated highly effective

adjuvant, developed by Jules Freund.
• It contains heat-killed Mycobacteria as an additional ingredient.
• Muramyl dipeptide, a component of the mycobacterial cell wall, activates
macrophages, making it highly effective.
• Both the Freund`s adjuvants are also an example of water-in-oil adjuvant.

• Aluminum potassium sulfate (alum) prolongs the persistence of antigen. When an

antigen is mixed with alum, the salt precipitates the antigen.

• The alum precipitate also increases the size of the antigen, thus increasing the
likelihood of phagocytosis.

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 Table showing the list of common Adjuvants

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