Antigen

ANTIGEN

• An antigen is a molecule that induces an immune response in

the body and helps in production of antibody or activate cells.
• Also called as immunogen.
• Any substance that is foreign and doesn't belong to the host
and satisfies two distinct immunologic properties-
o Immunogenicity
o Antigenicity.

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 Origin of antigen

Exogenous antigens
• Antigens that have entered the body from the outside.
E.g. By inhalation , ingestion , or injection .

Endogenous antigens
• Antigens that have been generated within previously normal cells
as a result of normal cell metabolism , or because of viral or
intracellular bacterial infection
 Immunogenicity
• Ability of an antigen to induce immune response in the body
(both humoral and/or cell mediated).
o B cells + antigen → effector B cells (plasma cell) + memory
B cells
o T cells + antigen → effector T cells (helper T cell or
cytotoxic T cell) + memory T cells

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 Antigenicity (immunological reactivity)
• Ability of an antigen to combine specifically with the final
products antibodies and/or T cell-surface receptors.

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Hapten:
Is a sma ll molecule t hat ca n elicit an immune response only
when atta ched to a large ca rrier suc h as a protein; t he carrier
may be one t hat also does not elicit an immune response by
itself.
Epitope:
The portion of an antigen that recognize and bound by
antibody and also called antigenic deter minant.
 ANTIGEN AND HOST RELATIONSHIP

• Based on the antigen-host relationship, antigens can

be grouped into two groups:
o Self or auto antigens
o Non-self or foreign antigens

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 Self or auto antigens
• Belong to the host itself - not immunogenic.
• Hosts do not react to their own antigens by exhibiting a
mechanism called immunological tolerance.
• Sometimes, the self-antigens are biologically altered (e.g. as
in cancer cells) and can become immunogenic.

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 Non-self or foreign antigens-

• Immunogenic and are of three types based on their phylogenetic

distance to the host.
o Alloantigens are species specific. Tissues of all individuals in a
species contain species-specific antigens.
o Isoantigens are type of antigens which are present only in subsets
of a species, e.g. blood group antigens and histocompatibility
antigens.
o Heteroantigens – Antigens belonging to two different species are
called as heteroantigens, e.g. antigens of plant or animal or
microorganisms etc. 9
 FACTORS INFLUENCING IMMUNOGENICITY

• Size of the antigen

• Chemical nature of the antigen
• Susceptibility of antigen to tissue enzymes
• Foreignness to the host
• Genetic factor
• Optimal dose of antigen
• Route of antigen administration:
• Repeated Number of doses of antigens
• Multiple antigens:
• Effect of prior administration of antibody:
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 Size of the antigen
• Larger is the size; more potent is the molecule as an immunogen.

Chemical nature of the antigen

• Proteins are stronger immunogens than carbohydrates
followed by lipid and nucleic acids.

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 Susceptibility of antigen to tissue enzymes
• Only substances that are susceptible to the action of tissue
enzymes are immunogenic as degradation breaks antigens
and expose epitopes.

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 Foreignness to the host
• Higher is the phylogenetic distance between the antigen
and the host; more is the immunogenicity.

Optimal dose of antigen

• An antigen is immunologically active only in the optimal dose
range.

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 Route of antigen administration
• Immune response is better induced following parenteral
administration of an antigen.
• Depends on the type of antibody produced.

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 Repeated doses of antigens
• Repeated doses of antigens over a period of time are needed
to generate an adequate immune response.
• This is due to the role of memory cells in secondary immune
response.

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 BIOLOGICAL CLASSES OF ANTIGENS

• Depending on the mechanisms of inducing antibody

formation, antigens are classified as:
o T cell dependent (TD) antigens.
o T cell independent (TI) antigens.

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 T-dependent (TD) Antigens

• If antigens are processed and presented by antigen-presenting

cells (APCs) to T cells which leads to T cell activation.
• Activated T cells secrete cytokines that in turn stimulate the B
cells to produce antibodies.

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 T-independent (TI) Antigens

• Antigens such as bacterial capsule, flagella and LPS

(lipopolysaccharide) that do not need the help of T cells and APCs.
• Directly bind to receptors present on B cells and stimulate B cells.
• Increased secretion of non- specific antibodies.
• TI antigens can activate both mature and immature B cells. B cells
can only differentiate into activated cells.
• No memory cells formation.

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 Differences between T cell dependent and T cell independent antigens

T Independent Antigen T dependent Antigen

Structurally simple- LPS, Structurally complex- protein in
capsular polysaccharide, flagella nature

Dose dependent Immunogenic over wide range of

Immunogenicity dose
No memory Memory present
No antigen processing Antigen processing step is needed

Slowly metabolized Rapidly metabolized

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 Differences between T cell dependent and T cell independent antigens

T Independent Antigen T dependent Antigen

Activate B cells polyclonally Activate B cells monoclonally

Activate both mature and immature B Activate mature B cells only

cells
B cells stimulated against T independent B cells stimulated against T dependent
antigen do not undergo- antigen undergo
 Affinity maturation  Affinity maturation
 Class switch over  Class switch over
Antibody response is restricted to IgM Antibodies of all classes can be produced
and IgG3

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 Superantigens

• Superantigens are the third variety of biological class of

antigens, Unique feature of superantigens is, they can activate
T cells directly without being processed by antigen presenting
cells (APCs).

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Superantigens

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 Superantigen
Bacterial superantigen
Staphylococcal toxin-
 Toxic shock syndrome toxin-1(TSST-1); Exfoliative toxin;Enterotoxins
Streptococcal toxin- Streptococcal pyrogenic exotoxin (SPE)-A and C
Mycoplasma arthritidis mitogen-I
Yersinia enterocolitica
Yersinia pseudotuberculosis
Viral superantigen
Epstein-Barr virus associated superantigen
Cytomegalovirus associated superantigen
Rabies nucleocapsid
HIV encoded superantigen (nef- negative regulatory factor)
Fungal superantigen
Malassezia furfur

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Antibody
ANTIBODY

• Humoral basis of immunity

• Secreted by plasma cell
• Present on B cell membrane
• React with antigen - specifically
observable
 Antibody or immunoglobulin
• Specialized glycoprotein, produced from activated B cells
(plasma cells) in response to an antigen.
• Capable of combining with the antigen that triggered its
production.
• By electrophoresis, the serum proteins are separated into four
fragments- albumin, globulin α, β and γ.
• Antibodies are located in the γ-globulin fraction; because they
immunologically react with the antigen; they were given the
name as immunoglobulin.
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 Antibody or immunoglobulin
• Both the terms, immunoglobulin (Ig) and antibody are used
interchangeably; representing the physiological &functional
properties of same molecule respectively.
• Immunoglobulin (Ig) constitutes 20-25 per cent of total serum
proteins.
• There are five classes (or isotypes) of immunoglobulins
recognised-IgG, IgA, IgM, IgD and IgE.

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 STRUCTURE OF ANTIBODY

• An antibody molecule is a ‘Y-shaped’

heterodimer; composed of four
polypeptide chains.
• Two identical light (L) chains, and Two
identical heavy (H).
• Each H and L chain comprises of two
regions- variable and constant region and
this is called Two Fab and One Fc

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ANTIBODY STRUCTURE
 STRUCTURE OF ANTIBODY (cont..)

• H and L chain:
o All four H and L chains are bound to
each other by disulfide bonds, and
by noncovalent interactions such as
salt linkages, hydrogen bonds, and
hydrophobic bonds.
o All the chains have two ends- an
amino terminal end (NH3) and a
carboxyl terminal end (COOH).
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 STRUCTURE OF ANTIBODY (cont..)

• There are five classes of H chains and two classes of light

chains. Immunoglobulin class Heavy chain type
IgG γ(gamma)
IgA α (alpha)
IgM µ(mu)
IgD δ(delta)
IgE ε(epsilon)

• L chains are of two types- kappa (κ) and lambda (λ).

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 Variable and Hyper variable region

• Represents the antigen binding site of

the antibody.
• Within the variable region, there are
some zones (hot spots) that show
relatively higher variability in the amino
acid sequences.
• Called as hypervariable regions or
complementarity determining regions
(CDRs).
• Form the antigen-binding site. 32
 Constant regions

• Constant region
o Constitutes the remaining part of an Ig
molecule other than that of variable
region.
o The amino acid sequence of constant
region shows uniform pattern.
o A single antibody molecule has two
identical heavy chains and two identical
light chains; H2L2.
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 FUNCTIONS OF IMMUNOGLOBULINS

• Antigen binding (by Fab region)

o Protection of the host.
o Interaction with the antigen.
o Valency of an antibody refers to the number of Fab regions it
possesses. Thus, a simple monomeric antibody molecule has
a valency of two.

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 IMMUNOGLOBULIN CLASSES

• Based on five types of heavy chains, there are five classes

of immunoglobulins (lgG, IgA, IgM, IgD and IgE).

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 Immunoglobulin G (IgG)

• Constitutes about 70-80% of total Igs of the body.

• IgG has maximum daily production.
• Longest half-life of 23 days.
• Highest serum concentration.
• Level raised - chronic malaria, kala azar,
myeloma, 20 infections.
• IgG has four subclasses- IgG1, IgG2, IgG3
and IgG4
• Called General purpose antibody 36
 Functions of IgG

• IgG can cross placenta - hence provide immunity to the fetus

and new born- Natural passive immunity
• Complement fixing and Phagocytosis
• Mediates precipitation and neutralization reactions.
• IgG plays a major role in neutralization of toxins as it can easily
diffuse into extravascular space.
• IgG is raised after long time following infection and represents
chronic or past infection (recovery).
• Coagglutination
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 Immunoglobulin M (IgM)

• Among all Igs, IgM has highest molecular weight due to polymer of
five peptide subunits, and hence called millionaire molecule.
• Present only in intravascular compartment, not in body fluids or
secretions.
• 5-8% of serum immunoglobulin
• Half-life of 5 days
• lgM – Earliest immunoglobulin to be synthesised

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 Immunoglobulin M (IgM)

• IgM exists in both monomeric and

pentameric forms:
o When present as membrane-bound
antibody on B cells, it exists in
monomeric form.
o When present in secreted form, it is
pentameric in nature

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 Functions of IgM

• Presence in serum - recent infection or Acute infection

• Complement fixing and Acts as an opsonin
• Protection against intravascular organisms
• Mediate agglutination
• Not transported across placenta
• lgM in fetus/newborn - indicates intrauterine infections-
Fetal immunity
• Short lived, disappear earlier than lgG
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 Immunoglobulin A (IgA)

• IgA is the second most abundant class of Ig next to IgG, constituting

about 10-15% of total serum Ig.
• Half-life of 6-8 days
• Major immunoglobulin – colostrum, saliva, tears
• Exists in both monomeric (Serum IgA) and dimeric forms (Secretory
IgA).
• IgA exists in two isotypes:
o IgA1
o IgA2
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 Serum IgA and Secretory IgA

• Secretory IgA; two IgA monomeric units joined

by a J chain.
• Secretory component Location-Predominant
antibody found in body secretions like milk,
saliva, tears, intestinal & respiratory tract
mucosal secretions.
• Dimeric secretory lgA - synthesised by plasma
cells, mucosal/glandular epithelial cells
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 Function of secretory IgA
• Role in local immunity or mucosal immunity against respiratory and
intestinal pathogens
• Resistant to digestive enzymes
• Inhibit adherence of organism to mucosa activates - alternate
complement pathway
• Promotes phagocytosis and intracellular killing of microorganisms
• Effective against bacteria like Salmonella, Vibrio, Neisseria, and viruses
like polio and influenza.
• Breast milk is rich in secretory IgA and provides good protection to the
immunologically immature infant gut.
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 Immunoglobulin E (IgE)

• Lowest serum concentration.

• Shortest half life.
• Minimum daily production.
• Only heat labile antibody (inactivated at 56º C in one hour).
• Has affinity for the surface of tissue cells (mainly mast cells)
of the same species (homocytotropism).
• Extravascular in distribution.
• Mediator of type I hypersensitivity reactions
• IgE is elevated in helminthic infections. 44
 Immunoglobulin D (IgD)

• IgD is found as membrane Ig on the surface of inactivated B cells

and acts as a B cell receptor along with IgM.
• Has the highest carbohydrate content among all the Igs.
• Resembles lgG structurally
• Concentration-3 mg/100 ml serum intravascular
• No other function is known for IgD so far.

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 Properties of various immunoglobulins

Property IgG IgA IgM IgD IgE

Usual form Monomer Monomer,dimer Monomer,Penta Monomer Monomer
mer
Valency 2 2 or 4 2 or 10 2 2
Other chains None J chain, J chain None None
secretory
component
Subclasses G1, G2, G3, G4 A1, A2 None None None
Molecular weight (kDa) 150 150-600 900 150 190
Serum level mg/mL 9.5–12.5 IgA1- 3.0 1.5 0.03 0.0003
IgA2 - 0.5

% of total serum Ig 75–85% 10–15% 5–10% 0.3% 0.019%

Half-life, days 23* 6 5 3 2.5
Daily production mg/kg 34 24 3.3 0.4 0.0023

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 Properties of various immunoglobulins

Property IgG IgA IgM IgD IgE

Intravascular distribution 45% 42% 80% 75% 50%
(%)
Sedimentation coefficient 7 7 19 7 8
Complement activation
Classical ++ (IgG3>1>2) – +++ – –
Alternate - + - - -
Binds to Fc receptors of ++ - ? ** - -
phagocytes
Placental transfer Yes (except IgG2) - - - -
Mediates coagglutination Yes (except IgG3) - - - -
Mucosal transport - Yes - - -
Mast cell degranulation - - - - yes
Marker for B cells - - + + -
Heat stability + + + + -
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 ABNORMAL IMMUNOGLOBULINS

Bence Jones • Produced in a neoplastic condition of plasma cells called multiple myeloma
proteins • Cancerous plasma cells produce excess of light chain (Bence Jones proteins) which
are accumulated in patient’s serum and excreted in urine.
Waldenstrom’s • Lymphoma affecting B cells producing excess IgM. It has been seen in multiple
macroglobulinemi myeloma.
a
Heavy chain • Characterized by an excessive production of heavy chains that are short and
disease truncated.
• Four types of heavy chain disease have been recognized based on H chain involved-
alpha chain disease (Seligmann's disease); gamma chain disease (Franklin's disease);
mu chain disease;delta chain disease.
Cryoglobulinemia • A type of Ig that becomes insoluble (precipitate) at low temperatures but redissolves
again if the blood is heated.
• Cryoglobulins have been associated with multiple myeloma and hepatitis C infection.

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