TOPIC: Antigens Lecture #3

ANTIGENS

1) Definitions
a) Immunogen - a substance that induces a specific immune response.
b) Antigen (Ag) - a substance that reacts with the products of a specific immune response.
c) Hapten - a substance that is nonimmunogenic but which can react with the products of a specific
immune response. Haptens are small molecules which could never induce an immune response
when administered by themselves but which can when coupled to a carrier molecule. Free
haptens, however, can react with products of the immune response after such products have
been elicited. Haptens have the property of antigenicity but not immunogenicity.
d) Epitope or Antigenic Determinant - the portion of an antigen that combines with the products of
a specific immune response.
e) Antibody (Ab) - a specific protein which is produced in response to an immunogen and which
reacts with an antigen.

2) Factors influencing immunogenicity

a) Contribution of the immunogen: The immune system normally discriminates between self and non-self
such that only foreign molecules are immunogenic. There is not absolute size above which a substance will be
immunogenic. However, in general, the larger the molecule the more immunogenic it is likely to be. In general, the
more complex the substance is chemically the more immunogenic it will be. The antigenic determinants are created
by the primary sequence of residues in the polymer and/or by the secondary, tertiary or quaternary structure of the
molecule. The physical form, such as whether it is particulate or soluble can affect immunogenicity. In general,
particulate antigens are more immunogenic than soluble ones and denatured antigens more immunogenic than the
native form. Antigens that are easily degradable and phagocytosed are generally more immunogenic. This is
because for most antigens (T-dependant antigens, see below) the development of an immune response requires that
the antigen be phagocytosed, processed and presented to helper T cells by an antigen presenting cell (APC).
b) Contribution of the Biological System: Genetic factors of the host can determine immunogenicity of an antigen.
Some substances are immunogenic in one species but not in another. Similarly, some substances are immunogenic
in one individual but not in others (i.e. responders and non-responders). The species or individuals may lack or have
altered genes that code for the receptors for antigen on B cells and T cells or they may not have the appropriate
genes needed for the APC to present antigen to the helper T cells. Age can also influence immunogenicity. Usually
the very young and the very old have a diminished ability to mount an immune response in response to an
immunogen.

c) Method of Administration: The dose of administration of an immunogen can influence its immunogenicity.
There is a dose of antigen above or below which the immune response will not be optimal. For example, high doses
of antigen can often be tolerogenic and blunt the immune response. Generally the subcutaneous route is for
effective for inducing an immune response than the intravenous or intragastric routes. The route of antigen
administration can also alter the nature of the response. Substances that can enhance the immune response to an
immunogen are called adjuvants. The use of adjuvants, however, is often hampered by undesirable side effects
such as fever and inflammation.
2) Chemical nature of immunogens

a) The vast majority of immunogens are proteins. These may be pure proteins or they may
be glycoproteins or lipoproteins. In general, proteins are usually very good immunogens.
Pure polysaccharides and lipopolysaccharides are good immunogens. Nucleic acids are
usually poorly immunogenic. However they may become immunogenic when single
stranded or when complexed with proteins. In general lipids are non-immunogenic,
although they may be haptens. Some glycolipids and phospholipids can stimulate T cells
and produce a cell-mediated immune response.
1) Types of antigens

a) T-independent antigens are antigens which can directly stimulate the B cells to produce antibody
without the requirement for T cell help. In general, polysaccharides are T- independent antigens.
The responses to these antigens differ from the responses to other antigens. These antigens are
characterized by the same antigenic determinant repeated many times as illustrated in Figure 1a.
Many of these antigens can activate B cell clones specific for other antigens (polyclonal
activation). T-independent antigens can be subdivided into Type 1 and Type 2 based on their
ability to polyclonally activate B cells. Type 1 T-independent antigens are polyclonal activators
while Type 2 are not. T- independent antigens are generally more resistant to degradation and
thus they persist for longer periods of time and continue to stimulate the immune system.

b) T-dependent antigens are those that do not directly stimulate the production of antibody without
the help of T cells. Proteins are T-dependent antigens. Structurally these antigens are
characterized by a few copies of many different antigenic determinants.

c) Hapten-carrier conjugates are immunogenic molecules to which haptens have been covalently
attached. The immunogenic molecule is called the carrier. Structurally these conjugates are
characterized by having native antigenic determinants of the carrier as well as new determinants
created by the hapten (haptenic determinants) as illustrated in the Figure 1c. The actual
determinant created by the hapten consists of the hapten and a few of the adjacent residues,
although the antibody produced to the determinant will also react with free hapten. In such
conjugates the type of carrier determines whether the response will be T-independent or T-
dependent.
 Fig 1A. B. C.

3) Antigenic determinants recognized by B cells and Ab

a) Antigenic determinants recognized by B cells and the antibodies secreted by B cells are
created by the primary sequence of residues in the polymer (linear or sequence
determinants) and/or by the secondary, tertiary or quaternary structure of the molecule
(conformational determinants). In general antigenic determinants are small and are
limited to approximately 4-8 residues. (amino acids and or sugars). The combining site
of an antibody will accommodate an antigenic determinant of approximately 4-8 residues.
Although, in theory, each 4-8 residues can constitute a separate antigenic determinant, in

practice, the number of antigenic determinants per antigen is much lower than what
would theoretically be possible. Usually the antigenic determinants are limited to those
portions of the antigen that are accessible to antibodies as illustrated in the Figure 2
(antigenic determinants are indicated in black).
4) Determinants recognized by T cells

a) Antigenic determinants recognized by T cells are created by the primary sequence of

amino acids in proteins. T cells do not recognize polysaccharide or nucleic acid antigens.
This is why polysaccharides are generally T-independent antigens and proteins are
generally T-dependent antigens. The determinants need not be located on the exposed
surface of the antigen since recognition of the determinant by T cells requires that the
antigen be proteolytically degraded into smaller peptides. Free peptides are not
recognized by T cells, rather the peptides associate with molecules coded for by the
major histocompatibility complex (MHC) and it is the complex of MHC molecules +
peptide that is recognized by T cells. Some T cells can recognize lipids in conjunction
with a MHC-like molecule called CD1. In general antigenic determinants are small and
are limited to approximately 8-15 amino acids. Although, in theory, each 8-15 residues
can constitute a separate antigenic determinant, in practice, the number of antigenic
determinants per antigen is much less than what would theoretically be possible. The
antigenic determinants are limited to those portions of the antigen that can bind to MHC
molecules. This is why there can be differences in the responses of different individuals.

5) Superantigens

a) When the immune system encounters a conventional T-dependent antigen, only a small fraction
(1 in 104 -105) of the T cell population is able to recognize the antigen and become activated
(monoclonal/oligoclonal response). However, there are some antigens which polyclonally
activate a large fraction of the T cells (up to 25%). These antigens are called superantigens
(Figure 3). Examples of superantigens include: Staphylococcal enterotoxins (food poisoning),
Staphylococcal toxic shock toxin (toxic shock syndrome), Staphylococcal exfoliating toxins
(scalded skin syndrome) and Streptococcal pyrogenic
 exotoxins (shock). Although the bacterial superantigens are the best studied there are
superantigens associated with viruses and other microorganisms as well. The diseases
associated with exposure to superantigens are, in part, due to hyper activation of the
immune system and subsequent release of biologically active cytokines by activated T
cells.

Figure 3

What is B Cell Receptor?

The B cell receptor (BCR) is a transmembrane receptor protein located on the outer surface
of B cells. B cells are produced as well as mature in the bone marrow. The B cell development
is initiated by the production of a functional pre-B cell receptor (pre-BCR). The pre-BCR
consists of two heavy chains and two surrogate light chains. These chains cooperate with IgA
and IgB which are signaling molecules. The BCRs which is also known as integral membrane
proteins reside in many identical copies at the surface of the B cells.

The B cell receptor complex is composed of an antigen binding subunit (MIg) which is made of
two immunoglobulin heavy chains and two immunoglobulin light chains and a disulphide-
linked heterodimer of Ig-alpha and Ig–beta proteins together, that make up a signaling
subunit. The heavy chains of BCRs consist of gene segments like 51 VH, 25 DH, 6 JH and 9
CH. 51 VH segments that encode the N terminal of the antibody. This N terminal of the
antibody includes the first two hyper-variable regions. 25 DH segment is a diversity gene
segment which encodes the third part of the hyper-variable region. 6 JH is the joining gene
segment which encodes the V region, and the 9 CH segment encodes the C region of the
BCR.
BCRs have a specific binding site, and this site binds to a region of the antigen called the
antigenic determinant. The binding is aided by non-covalent forces, the complementarity of
the receptor surface and the surface of the antigenic determinant. If the BCR is present on the
surface of B lymphocytes, it transmits intracellular signals which help in the regulation of cell
growth and differentiation while also binding to specific antigens to generate an immune
response. Memory cells that move through the circulation to produce immune responses are
also produced by the activation of BCRs. The antigens which are bound to this, occur with the
engulfment by the B cells due to receptor-mediated endocytosis. Then the antigens are being
digested into small fragments and are later displayed at the surface of the cells inside the
class II histocompatibility molecule.

What is T Cell Receptor?

T cell receptor (TCR) is found on the surface of T lymphocytes. TCRs function is to recognize
foreign particles known as antigens to initiate an immunological response. During normal
conditions, the body develops and produces many T cells, and each of the cells possesses a
unique TCR on its surface. The development of TCR occurs due to the recombination of
genes which encode TCRs prior to the encounter of antigens. In the surface of a T cell,
identical TCRs occur in larger quantities. The antigens which bind with the TCRs are small
peptide particles which are epitopes that occur through the phagocytosis of the foreign
pathogen. These epitopes are displayed by Major Histocompatibility Complex (MHC)
molecules.

T cells are of two types. Cytotoxic T cells (Tc) and Helper T cells (Th). The TCRs present on
Tc cells recognize foreign epitopes which are presented by MHC Class I molecules. They
possess the ability to differentiate nonself (foreign) antigens from self-antigens. Therefore, it
prevents the occurrence of immune responses against the body’s own cells. Th cells
recognize antigens displayed on MHC Class II molecules. A surface glycoprotein CD8 in Tc
cells and CD4 in Th involve during the binding process of the foreign epitope to both types of
T cells. CD4 and CD8 co-receptors recognize antigens presented on MHC Class II and MHC
Class I molecules respectively.
 Figure: T Cell Receptor
What are the Similarities Between B Cell Receptor and
T Cell Receptor?
 Both receptors are integral membrane proteins.
 Present on the cell surface as many identical copies.
 Both types possess unique binding sites.
 Both types of receptors are encoded by genes that are assembled through
recombination of segments of DNA.
 Both receptors bind to the antigenic determinant portion of the antigen, and the binding
occurs through noncovalent forces.

What is the Difference Between B Cell Receptor and T Cell Receptor?

B Cell Receptor vs T Cell Receptor

B cell receptor is a transmembrane T cell receptor is an antigen recognizing
receptor protein located on the outer surface molecule present on the surface of T
of B cells. lymphocytes.
Recognition of Epitope-antigens
B cell receptor recognizes soluble antigens. T cell receptor recognizes antigens displayed
on MHC Class I and MHC Class II molecules.