Antigen

Aditi Thakur- MSc Medical

Biochemistry-3rd year
06/20/2023 1
 Antigen
• Antigen is a substances usually protein in nature and sometimes
polysaccharide.

• Antigens are substances that react with antibodies.

• Immunogens are molecules that induce an immune response.

• Molecules that can be recognized by the immunoglobulin receptor of

B cell or by the T-cell receptor when complexed with major
histocompatibility complex (MHC) are called antigen.

• All antigens are not immunogens.

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 Epitopes
• An epitope is defined as the immunologically active
region of an immunogen.
• Consisting of 4 or 5 amino acids or monosachharide
residues.
• Binds to antigen-specific membrane receptors on
lymphocytes or secreted antibodies known as paratope.

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 Antigen with epitopes

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 Types of epitopes
• Sequential/ linear- Epitopes may be present as a single linear
sequence of primary configuration.
• Recognized by T cells.
• Non-sequential/conformational- Epitopes are found on the flexible
region of a tertiary antigen formed by bringing together the
surface residues from different sites of peptide chain during
folding.
• Recognized by B cells.

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 Cont…

• Linear epitopes on digestion of antigen

maintain the sequence.
• On digestion of antigen the conformational
epitope loses their sequence.

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 Linear and conformational epitope

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 Haptens

• Antigens that are not immunogenic but can

take part in immune reactions are termed as
haptens.
• Not immunogenic but antigenic.

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 Haptens
• Haptens are small organic molecules that are antigenic but not

immunogenic.

• They are not immunogenic because they cannot activate helper T

cells.

• Due to their inability to bind to MHC proteins.

• Haptens are univalent cannot activate B cells by themselves.

• Can activate B cells when covalently bound to a “carrier” protein.

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 Cont…
• When bound with a carrier molecule, they form an immunogenic hapten–

carrier conjugate.

• In this process, the haptens combine with an IgM receptor on the B cells,

and hapten–carrier protein complex is internalized.

• Peptide of the carrier protein is presented in association with class II MHC

protein to the helper T cells.

• Activated helper T cells produce interleukins which stimulate the B cells

to produce antibodies to haptens.

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 Cont…

• Hapten–carrier molecule is then taken in, processed,

and pieces of the carrier are presented by these B
cells and T h cells.
• In the body, the formation of hapten–carrier
conjugates is the basis for development of allergic
responses to drugs, such as penicillin.

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 Hapten–carrier conjugate

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 Immunogenicity & antigenicity
• Immunogenicity means the ability of an antigen to elicit an
immune reaction in the form of B-cell or T-cell response.

• Antigenicity means the ability to combine specifically with

the products of the above responses.

• All molecules that are immunogenic are antigenic but all

antigenic molecules cannot be considered immunogenic.

• Haptens lack immunogenicity.

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 Chemical nature of antigens

• Proteins- may be glycoproteins or lipoproteins.

• Polysaccharides- polysaccharides and lipopolysaccharides. 

• Nucleic Acids- nucleic acids are poorly immunogenic.

• They may become immunogenic when single stranded or

when complexed with proteins.

• Lipids- lipids are non-immunogenic, they may be haptens.

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 Types of antigen

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 Types of antigen on the basis of order of
their class (origin)

Exogenous Tissue Microbial

antigens
antigen antigen

Endogenous
antigens-Allogenic/iso/homologous,
xenogenic/heterophile, autologous/self.

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 Microbial antigens

Somatic Flagellar Capsular

O H K

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 Flagellar antigens
• Heat-labile proteins.

• Mixing salmonella cells with flagella-specific antisera

gives a characteristic pattern of agglutination.
• Bacteria are loosely attached to each other by their flagella
and can be dissociated by shaking.
• Antiflagellar antibodies can immobilize bacteria with
corresponding H antigens.

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 Capsular polysaccharide (CPS) / K antigen

• Associated with cell surface non-covalently and protects

bacteria from host immune system.

• K antigen repeat units form a high molecular weight structure

called as 'capsule'.

• Antigen serotyping is useful in understanding the pathogenesis

and for epidemiological studies in tracing the origin of disease.

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 Functions of capsules
• Prevention of dessication.

• Adherence.

• Resistance to non-specific host immunity (complement and complement-mediated

opsonophagocytosis).

• Resistance to specific host immunity (Because these capsules are structurally similar

to polysaccharides encounteres on host tissue, these capsules are poorly

immunogenic, and elicit a poor antibody response in the host.).

• Mediating the diffusion of molecules through the cell surface (The microbial proteins

synthesized are transported outside the cell).

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 Somatic antigen

• Antigen located in the cell wall of a bacterium

in contrast to one in the flagella (flagellar
antigen) or in a capsule (capsular antigen).

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 Soluble antigen

• Antigen dissolved in a liquid.

• A soluble antigen is recognized by B

lymphocytes but cannot be detected by T
lymphocytes until it has been processed by an
antigen-presenting cell. 

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 Exogenous antigens

• Antigens enters the body or system and start

circulating in the body fluids and trapped by the
APCs (Antigen processing cells such as
macrophages, dendritic cells, etc.).
• The uptakes of these exogenous antigens by APCs are
mainly mediated by the phagocytosis.
• Examples: bacteria, viruses, fungi etc
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 Endogenous antigens

• These are body’s own cells or sub fragments or

compounds or the antigenic products that are

produced.

• The endogenous antigens are processed by the

macrophages which are later accepted by the cytotoxic

T – cells.
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 Cont…

• Endogenous antigens include xenogenic

(heterologous), autologous and idiotypic or

allogenic (homologous) antigens.

• Examples: Blood group antigens, HLA

(Histocompatibility Leukocyte antigens), etc.

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 Autoantigens

• An autoantigen is usually a normal protein or complex of proteins (and

sometimes DNA or RNA).

• Recognized by the immune system of patients suffering from a specific

autoimmune disease.

• Due to genetic and environmental factors.

• Normal immunological tolerance for such an antigen has been lost in

these patients.

• Examples: Nucleoproteins, Nucleic acids, etc.

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 Types of antigens on the basis of immune response

• Complete Antigen or Immunogen

• Incomplete Antigen or Hapten

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 Complete antigen or immunogen

• Posses antigenic properties- able to generate

an immune response by themselves.
• High molecular weight (more than 10,000).
• May be proteins or polysaccharides.

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  Incomplete antigen or hapten

• These are the foreign substance, usually non-protein substances.

• Unable to induce an immune response by itself, they require

carrier molecule to act as a complete antigen.

• The carrier molecule is a non-antigenic component and helps in

provoking the immune response.

• Example: Serum Protein such as albumin or globulin.

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 Cont…

• Low Molecular Weight (Less than 10,000).

• Haptens can react specifically with its corresponding

antibody.

• Examples: Capsular polysaccharide of pneumococcus,

polysaccharide “C” of beta haemolytic streptococci,

cardiolipin antigens, etc.

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  Incomplete Antigen or Hapten

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 Determinants of antigenicity

• Whole antigen does not evoke immune response and

only a small part of it induces B and T cell response.

• Small area of chemical grouping on the antigen

molecule that determines specific immune response

and reacts specifically with antibody is called

an antigenic determinant. 
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Important determinants of antigenicity

• A number of factors have been identified that make a

substance immunogenic.
• Molecular size
• Foreignness
• Chemical-structural complexity

• Stability
• Other factors
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 Molecular size
• Protein molecules with large molecular weight are highly antigenic.

• Substances with molecular weights of about 100,000 Da and more

are highly immunogenic.

• Substances with molecular weights of less than 5000 Da are not

immunogenic.

• Substances with low molecular weight may be made antigenic by

adsorbing these on carrier particles, such as bentonite, kaolin.

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 Foreignness
• To be immunogenic- a molecule must be recognized as nonself, i.e.,
Foreign

• Foreignness implies ability of the host to tolerate selfantigens.

• A graft from an unrelated human will be rejected within about 2 weeks

unless immunosuppressive drugs are used.

• A graft from a chimpanzee will be rejected within hours even if drugs are
used.

• In contrast, a kidney graft from an identical twin will be accepted readily.

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 Chemical-structural complexity
• Proteins are the most potent immunogens followed by

polysaccharides.

• Nucleic acids and lipids are not efficient in eliciting a good immune

reaction- they may act as haptens.

• Structural complexity of a protein contributes to its immunogenicity.

• Chains of single amino acids or single sugars are poorly

immunogenic.

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 Cont…

• If different amino acids or sugars are combined in the

same molecule, the immunogenicity is greatly enhanced.

• Antibodies are produced by treating lipids with haptens

and then conjugating with suitable carrier molecules,

such as the proteins (e.g., hemocyanin or bovine serum

albumin).

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 Stability
• Highly stable and non-degradable substances (e.g. plastics, metals, or

chains of amino acids) are not immunogenic.

• Because internalization, processing, and presentation by antigen-

presenting cells (APCs) are essential to mount an immune response.

• Very stable substances (such as silicon) have been successful as non-

immunogenic materials for reconstructive surgeries, such as breast

implants.

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 Cont…
• If a substance is very unstable, it may break up before it can be

internalized, and hence become immunogenic.

• Large, insoluble complexes are more immunogenic than smaller,

soluble ones.

• Because macrophages find it easier to phagocytose, degrade,

and present the insoluble complexes than the soluble complexes.

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 Other Factors
• Biological system-

• Role in determining the immunological efficiency of an antigen.

• Substances are immunogenic in one individual but not in others (i.e.,

responders and non-responders).

• This is due to the fact that individuals may lack or have altered genes
that code for the receptors for antigen on B cells and T cells.

• They may not have the appropriate genes needed for the APC to
present antigen to the helper T (TH) cells.

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 Dosage and route of antigen
• Very low doses of antigen do not stimulate immune response.

• Because few lymphocytes are contacted or because a nonresponsive state is

elicited.

• An extremely high dose also fails to elicit tolerance

• Repeated administration of antigens (booster doses) may be required to enhance

immune response of the host to certain antigens.

• Booster doses of vaccines, such as DPT (diphtheria, pertussis, tetanus), DT

(diphtheria, tetanus), etc. are given to ensure good protective levels of
antibodies.

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 Adjuvants
• Substances- when mixed with an antigen and injected with it
boost the immunogenicity of the antigen.
• Increase both the strength and the duration of immune response.
• Boost immunogenicity of antigens.
• Aluminum potassium sulfate (alum) and freund’s water-in-oil
adjuvant prolong the persistence of antigen by forming a depot
at the injection site.

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 Antigenic Specificity

• Antigenic specificity of the antigen depends

on antigenic determinants or epitopes.
• B-cell epitopes
• T-cell epitopes

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 B-cell epitope
• Antigenic determinants recognized by B cells.
• B-cell epitope can combine with its receptor only if the
antigen molecule is in its native state.
• About six or seven sugar residues or amino acids long.
• Hydrophilic.
• Located at bends in the protein structure.

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 T-cell epitope

• T cells recognize amino acids in proteins but do not

recognize polysaccharide or nucleic acid antigens.
• Polysaccharides are considered as T-independent
antigens and proteins as T-dependent antigens.
• T-cell epitopes or antigenic determinants are small
and are only 8–15 amino acids long.

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 Species Specificity

• Tissues of all individuals in a species possess

certain species specific antigens.
• Some degree of cross-reaction occurs between
antigens from related species.
• Species specificity shows phylogenetic
relationship.
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 Phylogenetic relationship is useful in-

• Tracing evolutionary relationship between

species.
• Species identification from blood and seminal
stains in forensic medicine.

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 Isoantigens
• Isoantigens are antigens found in some, but not all, members
of a species.

• Species may be grouped depending on the presence of

different isoantigens in its members.

• Human erythrocyte antigens, based on which individuals are

classified into different blood groups, are the best examples
of isoantigens in humans.
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 ABO blood grouping system

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Blood groups are of primary importance in

• Transfusion of blood and blood products.

• Isoimmunization during pregnancy, and

providing valuable evidence in paternity
disputes.
• Results of which are supplemented by more
recent DNA fingerprinting tests.
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 Isospecificity

• Isospecificity is determined by the presence of

isoantigens or histocompatibility antigens.

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 Histocompatibility Antigens
• Histocompatibility antigens are the cellular determinants specific

for each individual of a species.

• Antigens are associated with the plasma membrane of tissue cells.

• Human leukocyte antigen (HLA) is the major histocompatibility

antigen that determines the homograft rejection.

• HLA typing is essential before carrying out transplantation of

tissue or organ from one individual to another.

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 Autospecificity
• Self-antigens are nonantigenic.

• Sequestrated antigens (such as eye lens protein and sperm) are

exceptions, because these are not recognized as self antigens.

• Corneal tissue and sperm are never encountered by the immune

system during the development of tolerance to self-antigens.

• These tissues become immunogenic if accidentally or

experimentally released into the blood or tissues.

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 Organ specificity
• Antigens characteristic of an organ or tissues are called organ-specific antigens.

• Found in brain, kidney, and lens tissues, even of different animal species, share the

same antigen specificity.

• Organ-specific antigens such as brain-specific antigens, shared by human and

sheep brain are one such example.

• Antirabies vaccines prepared from sheep brain, when given, may induce immune

response in some humans, causing damage to neural tissues of the recipient.

• May result in neuroparalytic complications in some individuals.

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 Heterophile specificity
• Same or closely related antigens, sometimes present in tissues of different
biological species, classes, or kingdoms are known as heterophile antigens.

• Heterophile specificity is determined by the presence of heterophile

antigens.

• Antibodies against the heterophile antigens produced by one of the species

cross-react with the antigens of other species.

• Weil–felix reaction, paul-bunnell test, and cold agglutination tests are the
examples of serological tests that use such heterophile antigens.

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 Superantigens
• Superantigens are a class of molecules that can interact with APCs and T
lymphocytes in a nonspecific way.

• Act differently by interacting with MHC class II molecules of the APC and the
Vb domain of the T-lymphocyte receptor.

• Results in the activation of a larger number of T cells (10%) than conventional

antigens (1%), leading to massive cytokine expression and
immunomodulation.

• Examples of superantigens are staphylococcal enterotoxins, toxic shock

syndrome toxin, exfoliative toxins, and also some viral proteins.

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 Superantigens

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 Reference

• Textbook of Microbiology & Immunology,

Second Edition- Subhash Parija.

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