Antigen

Introduction

Antigen is a foreign macromolecule, which can initiate an immune response inside

any host. Any large macromolecule can serve as an antigen including those in
bacteria, viruses, protozoa, helminthes foods, snake venom, serum components, red
blood cellsor cells and tissues of various species.

The antigen structure does not match any structure of the host (self), therefore
antigen in majority of cases is a non-self structure. B cells or T cells present in the
host identify these non-self structures as foreign, and mount immune response
against it.

Our immune system has designed various mechanisms for protecting our body from
diseases. The immune system requires some hint from the invading pathogen to
mount response against it. Cells of immune system recognize the invading pathogens
by the specific non-self structures present on them. These non-self structures are
some macromolecules present on invading bacteria, virus, fungi or any other parasite
which are known as Antigens. As soon as the presence of non-self structure is
registered by the host’s body, immune response is mounted against it. As this non
self structure leads to the generation of immune response, it is also known as an
Immunogen.

An antigen is basically a marker of any invading agent for a host cell, which can be
used to identify its foreignness. These antigens are a part of the basic structure of
any organism. These are the integral structures of the organism in which they are
present. These structures will not be considered as antigens for the host but they will
act as antigens for any other organism.

Every organism possesses some structures inside the cells or on the surface, which
can be identified as markers or antigens on these cells. Blood group antigens in
human beings, feather antigens in birds, cat hair antigens in cats, sheep RBC
antigens in sheep and bacterial antigens are a few examples. The structure of
outermost part of bacterium may contain some motifs which can function as
antigens, which are unique to the microorganisms. They are-

Somatic   Antigens Antigens present on the cell wall of a Gram negative

bacterium are known as somatic antigens. They are designated as ‘O’ antigens.
Capsular  Antigens Antigens in the bacterial capsule are capsular antigens. They
are denoted as ‘K’ antigens.
Flagellar   antigens Antigens present on the flagellum of an organism are known
as flagellar  antigens. They are denoted as ‘H’ antigens.
Protein substances such as exotoxins   and   endotoxin produced by pathogenic
microorganisms are also antigenic.

The terms antigens and immunogens are sometimes used synonymously but there
is a slight difference between them.

Immuno—gen: - a macromolecule which can generate immune response.

Immunogens can: -

Ø Bind to the antibodies

Ø Generate the immune response

Antigens can:-

Ø Bind to the antibodies

Immunogenicity and Antigenicity

Immunogenicity is the ability of any macromolecule by the virtue of which it can

generate the immune response and also bind to the antibodies developed as a result
of the response; whereas antigenicity refers to binding of antigens to their
complementary structures, the antibodies.

Antigenic Determinants

An antigen is usually a complex macromolecule folded in a specific pattern.

Immune cells interact with the antigens at restricted portions on the molecule. These
differential small motifs can bind to different types of antibodies and activate
different clones of B cells or T cells. These structural subunits are the sites where the
antibody actually binds an antigen and are known as Epitopes or Antigenic
Determinants, which bind to antibody's Paratope or Complementarity
Determining Regions. Antigens may possess more than one type of epitope. They
can have multiple epitopes of same type as well as different types.

The epitopes of antigens identified by T cells and B cells are different. The
epitopes identified by B cells are usually on the surface of antigens and may be
three-dimensional. T cells recognize only those peptides, which are exhibited, on
self MHC (Major Histocompatibility Complex) molecules. These peptides may be
from the embedded portions of protein antigens, which are exposed only after
processing or cleavage.

Exogenous and Endogenous Antigens

Exogenous antigens are the antigens which enter our body from external
sources.They enter our body during various infections, such as bacteria, viruses,
fungi, protozoa, helminthes etc., or environmental substances such as dust mites,
foodstuffs, pollens and pollutants etc. by inhalation, ingestion, or injection. The
immune response developed against these antigens protects us from their adverse
effects. The endogenous antigens are internally generated molecules due to normal
cell metabolisms or due to intracellular bacterial or viral infections. These antigens
are presented on the cell surface associated with class I histocompatibility
complexMHC class I. Some examples for endogenous antigens include self-
antigens, tumor antigens, alloantigens, and some viral antigens

Factors governing the immunogenicity of a macromolecule

Immunogenicity of any molecule depends on many factors. The possibility and

extent of immune response generation also depends upon the macromolecular
architecture. Properties governing the immunogenicity of any macromolecule are:-

Ø Molecular size
Ø Foreignness
Ø Chemical nature
Ø Heterogeneity
Ø Susceptibility to antigen processing and presentation
Molecular size

A macromolecule can generate an immune response if its molecular size is

high. Efficient immunogens are usually possess molecular weight above 10,000
Daltons. Macromolecules with high molecular size activate the B cells efficiently.
Small antigen molecules are unable to crosslink the B cell surface receptors,
therefore its stimulation is also prevented. Hemocyanin or serum albumin is more
immunogenic than insulin or glucagon.

Some low molecular weight macromolecules exhibit antigenecity but not

immunogenicity. They are known as Haptens. Haptens are the molecules, which can
bind to the antibodies but cannot elicit the immune response. They are low
molecular weight macromolecules, which cannot stimulate the B cells or T cells to
mount the immune response. Immune response can be generated against them by
binding these small size haptens to some carrier molecules. These carriers are
usually proteins. The antibodies developed against this conjugate may be against
hapten molecule, carrier molecule, and regions bridging hapten carrier complex.
Some drugs, peptide hormones and steroid hormones can function as haptens.

Foreignness

The immune response of our body depends on the degree of variability

between self and nonself structures. This variability is foreignness. The foreignness
increases with the increase in evolutionary difference. Immunogenicity increases
with increasing foreignness because the variation in macromolecular structures
increases. For example an immunogen from worms shall elicit potent immune
response in human beings as compared to an immunogen from goat. Goat is also a
vertebrate and possesses close evolutionary relationship with humans as compared to
worms. Microorganisms possess greater evolutionary divergence; therefore they
stimulate our immune cells with great efficiency.

Chemical Nature
 Chemical composition of molecules has an important role in contributing
towards their immunogenicity. Proteins are the most potent immunogens amongst all
biomolecules. After proteins, polysaccharides also serve as effective immunogens.
Cells of our immune system are less responsive towards lipids and nucleic
acids.Immune response to most of the infectious microorganisms istowards their
protein antigens. Proteins are highly immunogenic because of their large
macromolecular size and diversity in their structure.

Heterogeneity

Heterogeneity in the macromolecule enhances its immunogenicity.

Macromolecules composed of same repeating units are weekly immunogenic such as
homopolymersexample starch or cellulose. Natural heterogeneity exists to the
highest level in proteins, therefore majority of the antigens are proteins. Proteins
naturally contain different types of amino acids, which accounts for complexity in
the structure. Complexity further increases because of their primary, secondary,
tertiary and quaternary levels of structure. Heterogeneity in the macromolecular
structure further increases by complexing two types of biomolecules, for
instanceglycoproteinsand lipoproteins may be more immunogenic than proteins.
Immunogenicity of polysaccharides and lipids can be enhanced by conjugating them
with other biomolecules.

Susceptibility to Antigen Processing and Presentation

Generation of immune response against certain antigens requires their digestion

into smaller units. The proteins antigens which can be engulfed by antigen
presenting cells (APCs) are cleaved into polypeptides. These short stretches of
peptides can be presented to the T cells, which inturn will lead to immune response.
Antigens which cannot be digested and presented by antigen presenting cells are
poor immunogens.

Specificity

Specificity of an antigen is its property to bind to a specific antibody only and

not others. The antibody raised against an antigen binds to it by the complementary
structure. The binding will occur only if the attachment sites on antigen and antibody
exactly match each other. In very rare cases the antigen binds to an antibody whose
structure is close to its complementary structure, this feature is known as cross
reactivity.

The specificity or cross reactivity of any antigen is the attribute of structural

similarity or dissimilarity in the macromolecules involved. The specificity or the
property of an antigen binding to a specific antibody is due to the compatibility in
their binding sites. If two different antigens possess some bit of similarity in their
structure, they may bind to same antibody. This can be further understood by
classifying antigens into various categories.

Types of Antigens

I. Organ Specific Antigens

II. Species Specific Antigens
III. Autoantigens  
IV. Alloantigens  
V. Heterophile  antigens  

I. Organ Specific Antigens

Organ specific antigens are the antigens associated with a particular organ of all
the members of the same species. These antigens are associated with the given organ
and not found on other tissues. Eg. thyro- globulin is specific for thyroid. Structural
units of any given organ in all the members have same function; for example all the
nephrons of the kidney perform same function of blood filtration, therefore these
cells posses similar structure for performing the same specific functions. Nephrons
of all the members of the species will have same structure therefore same antigens.
This structural similarity accounts for presence of organ specific antigens. These
antigens can be the markers of that particular organ. It may also be present on that
particular organ in even unrelated species example adrenal antigens, kidney
antigens.

II. Species Specific Antigens

 These antigens are the markers of a particular species. They are present in all
the members of any given species and are not present in other species. These
antigens help in distinguishing the species immunologically for   example,   serum  
albumin of horses is immunologically  different  from  that of man,  dog,  sheep,  and
so on.  These  antigens  can  also  be  helpful  in  tracing  evolutionary  relationships.  

III. Autoantigens  

  Sometimes   the   immune   system   identifies   the   self-­‐structures   as   foreign   and  

mounts   an   immune   response   against   it,   these   structures   are   the   autoantigens.  
These  antigens  are  the  self-­‐antigens  of  any  host,  but  due  to  some  reasons  these  are  
identified   as   foreign   by   the   immune   system.   Cell   mediated   or   humoral   Immune  
response  is  mounted  against  these  antigens  which  lead  to  its  destruction  and  inturn  
the  destruction  of  self  tissues.  Recognition  of  self  structures  as  foreign  is  observed  
in  many  disordersnamed  as  autoimmune  disorders  for  example  Multiple sclerosis,
Myasthenia gravis, Pernicious anemia, Rheumatoid arthritis, etc.  

IV. Alloantigens  

  These  antigens  are  present  in  some  members  of  any  given  species.  The  cells  
of   some   members   of   a   given   species   carry   one   form   of   the   antigen   and   another  
carries   a   different   form   of   structure.   This   difference   is   due   to   the   presence   of  
different   alleles   in   different   individuals.   They   are   genetically   determined.   These  
antigens  can  be  used  to  categorize  the  members  of  the  species  for  example  blood  
group   antigens.   These   antigens   are   of   clinical   importance.   Major   Histocompatibility  
(MHC)   antigens   also   fall   under   this   category.   They   are   also   sometimes   termed   as  
Isoantigens.  

V. Heterophile  antigens  

These are the antigens of similar nature present on organs or tissues of

unrelated species. These antigens may be structurally closely related but may occur
in different classes or even kingdom eg. Forssmann antigenis a lipid-carbohydrate
complex widely distributed in humans, animals, birds, plants and bacteria etc. It is
present on red blood cells of sheep, horse, cat, mouse, chicken, kidney cells of
guinea pigs and Pneumococci. These heterophile antigens have useful applications
in diagnosis of diseases. Ex. Weil-Felix test for typhus fever where patients suffering
from rickettsial disease produce antibodies, which can agglutinate some non-motile
strains of Proteus and Paul Bunnel test for infectious mononucleosis where patients
suffering from infectious mononucleosis due to EBV produce antibodies, which
agglutinate sheep erythrocytes.

Biological Classes of Antigens

Immune response to antigens is mounted by cell mediated as well as humoral

immunity. Cell mediated immune response is generated by presenting peptides by
self cells on MHCs to T cells and humoral immune response is generated against
antigens by the stimulation of B cells and eventually the production of
antibodies.There are two mechanisms for antibody production through humoral
immunity. Antigens can be classified on the basis of the mechanism they follow to
generate immune response. Some antigens provoke the B cells directly to produce
antibodies and others require the help of T helper cells. We can classify the antigens
into two types on this basis:--

I. T cell independent antigens (TI)

II. T cell dependent antigens (TD)

I. T cell independent antigens (TI)

These antigens do not require the involvement of T cells for the production of
antibodies. These antigens have repeating units of epitopes, which bind to the B cell
receptors and stimulate them for example bacterial LPS (Lipopolysaccharides). As a
result of stimulation B cells proliferate and differentiate into plasma cells which
eventually produce antibodies. Some of the B cells are stored as memory B cells for
further protection against these antigens.

II. T cell dependent antigens (TD)

Some antigens require the help of T helper cells for the production of
antibodies. They are processed by antigen presenting cells (APCs) and presented to
T helper cells. The large sized protein antigens are engulfed by the APCs and
cleaved into small peptides. These peptides bind to MHC and are presented on the
surface of APCs. The T cell receptors (TCR) on the surface of T helper cells bind to
the foreign peptides on self MHC and get activated. These T helper cells stimulate
the B cells for production of antibodies. Such antigens are known as T cell
dependent antigens.

Super antigens

Super antigens are a special category of antigens, which stimulate a large

population of T cells nonspecifically. They are potent T cell mitogens, which means
that these molecules have enormous capacity to stimulate the multiplication of T
cells. These antigens do not stimulate the T cells against some specific antigen but
only initiate its nonspecific proliferation strongly. These activated T cells exhibit
massive immune response and produce large amount of cytokines. This nonspecific
exaggerated immune response is harmful to our body as it causes shock and multiple
organ failure. They can be of bacterial or viral origin for example exotoxins
produced by pathogenic Streptococci.

Role of Biological System in Immunogenicity

The host plays a major role in generation of immune response. The immune
response depends on the genetic make-up of the host, the dose of incoming antigen
and it’s route.

Genotype of the host

An antigen given to two different hosts generates different levels of immune

response and the antibody titer may vary greatly in both the individuals. This is
influenced by the host’s genotype. The cell mediated and antibody mediated immune
response depends on the recognition of antigen molecule. The antigens are
recognized by either B cell receptors or T cell receptors. The B cells identify the
antigens and produce antibodies against them. If the antigenic stimulus exhibits high
degree of foreignness the production of antibodies by B cells will be high. The T
cells are stimulated by the peptides presented by self-MHC. The degree of T cell
stimulation and activation depends on sequence of peptides presented by MHC,
which inturn decides the extent of immune response.
Dose

The immune response also depends on the dose and route of administration. If
the number of antigens entering the host is very high or very low, then
immunological unresponsiveness or tolerance develops. The efficient response also
requires repeated doses of antigens, which enhances the number of activated B and T
cells.

Route of administration

The route of administration of the antigen has a profound effect on the

generation of immune response. The antigen entering through intravenous route may
not serve to be immunogenic whereas its intradermal entry may elicit enormous
response.

Adjuvants

Adjuvant is derived from the word ‘adjuvare’ which means ‘to help’.Adjuvants
help in enhancing the immune response. They are administered along with
immunogens and are non-immunogenic. Adjuvants increase the immune response by
releasing the antigen slowly. They also stimulate the macrophages nonspecifically
and enhance inflammatory effects.The addition of adjuvant increases the effective
time of antigen exposure to immune system from few days to several weeks which
helps in enhancing the immunity to a great extent.

They can be organic compounds or can be of microbial origin. Freund’s

adjuvant is commonly used for effective antigen administration. Freund’s complete
adjuvant is a preparation, which contains heat-killed Mycobacteria and muramyl
peptide, a component of Mycobacterial cell wall. Freund’s incomplete adjuvant is
water-in-oil emulsion, containing antigens in aqueous solution, mineral oil and an
emulsifying agent. Aluminum potassium sulfate (alum) is also a commonly used
adjuvant.

Applications of Immunogenicity-Antigens used as vaccines

The property of antigens to generate immune response is applied for the

preparation of vaccines. Vaccines are killed or attenuated preparations of
microorganisms. The antigenic preparation without the presence of toxic
components of pathogens is developed to immunize the host against this pathogen.
There are numerous different types of antigens on the bacterial surfaces. A single
organism such as a bacterium may contain a variety of proteins, carbohydrates and
other materials that provoke immune responses.

Conclusion

Immunogens are foreign macromolecules, which can induce the immune

response of any host. All organisms possess some structures in their cells or on their
surfaces which can function as antigens. Immunogenicity of any macromolecule
depends on its size, chemical nature, complexity of the molecular structure and
degree of foreignness. Large macromolecules like proteins, glycoproteins and
lipoproteins are highly immunogenic because of the complex nature. Antibodies
bind to the antigens at particular site known as epitopes. Immune response inside an
organism is influenced by its dose, route of administration and genotype of the host.
Immunogenicity is applied for preparations of vaccines, which provide immunity to
the hosts.