ANTIGENS

Properties:
Antigens- may possess: immunogenicity, antigenicity, allerogenicity, or
tolerogenicity
Immunogenicity - Property that allows a substance to induce a detectable
immune response (humoral or cellular) when introduced into an animal. Such
substances are termed Immunogens.
Antigenicity - Property that allows a substance to combine specifically with
antibodies or TcR , whether or not they are immunogenic. Therefore, all
immunogens are antigens but not all antigens are immunogens.
Allerogenicity- Property that allows a substnace to induce an allergic response.
Such substances are termed allergens.
Tolerogenicity- Property that allows a substance to induce specific immunologic
non-responsiveness in either the humoral or cell-mediated branch. Such
substances are termed tolerogens.
Haptens - Low-molecular weight compounds including many drugs and
antibiotics, are non-immunogenic but when coupled to immunogenic proteins, the
resulting conjugates stimulate the production of antibodies which can bind to the
low-molecular weight component. Such molecules are termed
haptens.
Epitope - the part of an antigen that combines with a specific antibody or T cell
receptor. Previous term used was antigenic determinant.
Immunogens
For the induction of humoral immunity (antibody response), the most potent
immunogens are macromolecular proteins or
glycoproteins, but polysaccharides, synthetic peptides, and other synthetic
polymers such as polyvinylpyrrolidone are immunogenic under
appropriate conditions. Pure nucleic acids or lipids are not immunogenic but
antibodies which react with them can be induced by immunization with
nucleoproteins or lipoproteins.
In general, proteins serve as immunogens for T cell-mediated immunity. (Recall
that these proteins must be processed into peptides and the peptides must be
presented by an APC in association with MHC proteins.
Requirements for Immunogenicity
The requirements are somewhat dependent upon the experimental conditions

(mode of immunization, organism being immunized, sensitivity of detection

methods, etc.). However, certain conditions must be met in order for a molecule
to be immunogenic:
A. Foreignness: [Rabbit albumin not immunogenic in another rabbit but would be
immunogenic in a mouse.]
B. Molecular Size: [Certain minimum size is required for immunogenicity. The
most potent immunogens are macromolecular proteins with
molecular weights greater than 100,000. Substances <1000 are not usually
immunogenic]
C. Chemical Complexity: [Homopolymers consisting of repeating units of a single
amino acid are poor immunogens regardless of their size]
Co-polymers of 2 or 3 amino acids may be good immunogens. A co-polymer of
glutamic acid and lysine must be 30-40,000 to be immunogenic
Add tyrosine - reduce size limitation to 10-20,000
Add tyrosine and phenylalanine and reduce to, 4000
-- In general, immunogenicity increases with structural complexity.
Aromatic amino acids contribute more to immunogenicity than non-aromatic (ex:
tyrosine or phenylalanine).
D. Degradability: Macromolecules that cannot be degraded and processed by
APCs are poor immunogens. For example, polymers of D-amino acids are not
immunogenic. Proteolytic enzymes can only degrade proteins containing L-amino
acids.
E. Genetic constitution of the Animal: Immune response is under genetic control.
Individuals differ in their ability to respond to immunogens.
Animal may not have an appropriate Ig, may not have an appropriate TcR, or
may not have an appropriate MHC.
F. Method of Antigen Administration: Whether an antigen will induce an immune
response depends on the dose and the mode of administration.
Antigen can be delivered in a variety of ways: ip, sc, iv, im, id, etc. The repsonse
will vary depending upon the route chosen. In addition, too little antigen may not
stimulate a strong response, whereas too much antigen may stimulate tolerance
(specific non-responsiveness). The use of an adjuvant can also enhance the
immunogenicity of an antigen. Adjuvants (such as Freunds) prolong the
persistence of the antigen in the body, induce a strong inflammatory response
which can lead to granuloma formation, and can stimulate lymphocyte
proliferation.
T cells and B cells exhibit fundamental differences in epitope recognition.
B cell epitopes [epitopes recognized by membrane-bound Ig, or secreted Ig]
The binding of antigen to Ig involves weak non-covalent interactions so there
must be complementarity.
Properties of B cell epitopes:
1]Epitopes may be associated with soluble immunogens or particulate

immunogens.
2]Epitopes tend to be accessible and on the exposed surface of the immunogen.
3]Epitopes generally contain hydrophilic amino acids
4]Often the epitopes are found where the molecule bends or where there is a
high degree of segmental mobility [atomic mobility]
5]The epitope may consist of either sequential or non-sequential amino acids. If
non-sequential, the 3D conformation of the epitope if vital.
6] Complex proteins contain multiple overlapping epitopes
7] The size of a B cell epitope is determined by the size of the antigen binding
site on the antibody molecule. Conformationally determined epitopes
tend to require a larger epitope. Smaller ligands (carbohydrates, peptides,
haptens, etc.) tend to fit within a deep concave pocket or crevice. Larger
globuluar antigens tend to interact with Ig across a large planar face. Protrusions
on the antigen binding site would be complementary with
depressions on the epitope and vice versa. This type of interaction is obviously
highly dependent upon the 3D conformation of the globular antigen.
T cell epitopes [epitopes recognized by membrane-bound TcR only]
T cells recognize processed peptides associated with MHC on the surface of
APCs (Class II MHC) or altered self cells (Class I MHC). In other words, T cells
exhibit MHC RESTRICTED ANTIGEN RECOGNITION
CD4+ T cells are restricted to Class II MHC
CD8+ T cells are restricted to Class I MHC
More recent evidence suggests that a small population of T cells may possess
TcRs capable of recognizing lipids or glycolipids. Little is known about this type
of recognition and we will restrict our discussion to peptide epitopes.
1]The binding of the TcR to MHC + peptide represents a tri-molecular complex.
2]Only oligomeric peptides serve as epitopes. Epitopes which bind to Class I
MHC have an optimal size of 9 amino acids with a range from 8-11. Epitopes
which bind to Class II MHC have an optimal size range from 12-25 amino acids.
3) Epitopes are often internal and only exposed by processing within APCs or
target cells.
3] Antigen processing is required to generate the peptides that interact
specifically with MHC molecules.
4] T cell epitopes must have two binding regions. The region of the peptide
which binds to MHC is termed the agretope while the region which binds to the
TcR is termed the epitope.
5) Complex proteins may contain multiple, overlapping epitopes.
6 ) Immunodominant T cell epitopes are determined by the set of MHC molecules
which are expressed by an individual.
Experimentally it has been demonstrated that there is a correlation between the

ability of a peptide to bind to a particular MHC molecule and the T

cell response to that peptide.
MITOGENS
Mitogens are agents that are able to induce cell division (mitosis) in a high
percentage of T or B cells. This proliferation is described as polyclonal activation
(as opposed to clonal expansion following the specific encounter with a
conventional immunogen). There are T cell mitogens and B cell mitogens.
A number of common mitogens are lectins. Lectins are proteins which bind to
specific carbohydrate groups (moieties). They bind to
glycoproteins on the surface of the lymphocytes and cause activation.
Important...they do not act via conventional TcR-epitope or Ig-epitope
interactions.
Lectin Examples:
Con A - T cell mitogen
PHA - T cell mitogen
PWM - T and B cell mitogen
Another important mitogen which is NOT a lectin is LPS (lipopolysaccharide).
This polysaccharide component of the outer membrane of gram - bacteria is also
known as endotoxin. LPS is a very potent mitogen for B cells.
SUPERANTIGENS
Among the most potent T cell mitogens known!!
Bind differently to the TcR and MHC so a large # of T cells are activated. In some
cases as many as 1/5 T cells may be activated to proliferate and to
secrete cytokines. See figure in text for nature of interaction of superantigen +
TCR and MHC. Many microbial pathogens (including Streptococcus and
Staphylococcus) produce toxins which function as superantigens. The
symptoms of staphylococcal food poisoning are due to the massive T cell
proliferation and cytokine secretion which occurs in the GALT due to the
presence of staphylococcal enterotoxin in food. Toxic shock syndrome and Toxic
shock-like syndrome are both due to the action of bacterial superantigens.