Unit 2

Antigen and Antibody

Antigen
 A substance that can be recognized by an immunoglobulin receptor of B cells or by a T cell
receptor when complexed with MHC class II receptors.
 A compound that evokes an immune response by the body is called an antigen
 The ability of a compound to bind with an antibody (final product of cell mediated and humoral
immune response) is called antigenicity
 An immunogen is any agent capable of inducing an immune response and is immunogenic.
 All immunogens are antigens, but not all antigens are immunogens.
 Immunogenicity is the ability to induce, a humoral and cell mediated immune response.
 All molecules that have the property of immunogenicity also have the property of antigenicity
but the reverse is not true.

Factors Affecting Immunogenicity or Antigenicity

 Nature of immunogen contributes to immunogenicity

 These are some properties of an immunogen
 Foreignness: Should be foreign with respect to the host. The adaptive immune system
recognizes and eliminates foreign antigens.
 Size: Certain minimal molecular weight. Small compounds with weight <1000 Da are not
immunogenic. Those greater than 6000 Da are immunogenic.
 Chemical complexicity: Carbohydrates can be complexed with proteins as glycoproteins.
o For example, homopolymers and copolymers.
 Dosage and route of administration: Insufficient dose of immunogen may not stimulate
an immune response since it’s not enough to activate lymphocytes.
 Lipids as Antigens: Use of lipid proteins conjugates.
 Susceptibility of antigen processing and presentation
 Biological system contributes to immunogenicity
 Genotype of the recipient animal

Haptens

 These substances fail to induce immune responses in their native form

 They have low molecular weight and their chemical simplicity
 They are antigenic but not immunogenic
 These compounds become immunogenic when conjugated with high molecular weight as
complex carriers.
 It is capable of inducing an immune response.
 When haptens are conjugated with a carrier, an immune response can be induced against it.
Types of Antigens

Exogenous Antigens Endogenous Antigens

Antigens present outside the cell Antigens present inside the cell
Ex: drugs, pollen grains (allergens), pollutants Cancer cells possessing viral particles (viral
antigens)

 Isoantigens: These are antigens present in the body. Ex: ABO blood group on RBC cells
 Isoantibodies: Foreign antibodies binding to isoantigens. Ex: ABO blood group on RBC cells

Epitopes

 A part of the immunogen which binds to the antigen

 There are two types of epitopes,

B cell epitope T cell epitope

Present on the surface Processed and complexed with MHC class II
receptors
Made up of hydrophilic amino acids Made up of hydrophobic amino acids (exposed by
exocytosis)
Can adjust the size according to the length of Can adjust the size according to the length of
antigen binding site of antibody antigen binding site of antibody

 Paratopes: This is the region on the antibody which binds to the antigens.
 Heterophile Antigen: Antigens of similar nature, if not identical, that are present in different
tissues in different biological species. Usually different species have different antigen sets but
the heterophile antigens are shared by different species.

Cross Reactivity

 An immunologic reaction in which a particular antibody or t cell receptor reacts with two or
more antigens possessing the same epitope.
 When antibodies or cells with a specificity to one epitope bind weakly to another epitope.

Ex: Streptococcus mylogenes – M antigen (heart, liver)

 Antibody

 Antigen binding glycoproteins are synthesized by B cells each with different amino acids
and different antigen binding sites.
 They are the most abundant proteins present in the blood
 Secreted by plasma cells
Structure of an Antibody
 They are Y-shaped molecules with
two identical antigen binding sites
each on one arm
 Since they have two antigen
binding sites, they are termed as
bivalents (Fab fragments)
 Composed of 4 polypeptide chains
 It is a heterodimer
o Two identical light chains
(L)
o Two identical heavy chains
(H)
 Each light and heavy chain are
bound together by disulphide
bridges or non-covalent linkages
 The tail end is known as the Fc
region.
 There are 2 pairs of variable chains: light and heavy chains
 There are 4 pairs of constant chains both light and heavy chains.
 The hinge region is rich in proline and cysteine residues and is flexible
 The different types of antibodies are: IgM (pentamer), IgA (dimer), IgG, IgD and IgE.
 They are expressed as either secreted immunoglobulins or membrane bound immunoglobulins.
o Secreted immunoglobulin’s have hydrophilic amino acid sequence at carboxyl end
o Membrane bound immunoglobulins have three regions at their carboxyl end,
extracellular hydrophilic spacer sequence, hydrophobic transmembrane sequence and a
short cytoplasmic tail.

Types of Immunoglobulins

1. IgM
 First class of antibodies developed in a B cell
 Accounts for 5%-10% of total serum proteins
 Found on surface of mature B cells with IgD
 Monomeric IgM has a molecular weight of 180,000
 Secreted by plasma cells as a pentamer – two light chains and two heavy chains
  Joined by disulphide bonds between their Fc portions and by a polypeptide chain – J
chain
 Half-life is approx. 5 days
 First class of Ab to respond to any antigen
 First to be synthesized neonate
 More efficient than IgG in activating a
complement (requires two Fc regions in
close proximity)

2. IgA
 Constitutes 10%-15% of total serum Ab proteins
 Present predominantly in colostrum, saliva, tears, mucus of bronchi, genitourinary and
digestive tracts.
 Half-life is 5.5 days
 Two subclasses: IgA1 and IgA2
 Predominantly monomeric but external secretions of IgA (secretory IgA) are dimers or
tetramers
 Consist of a J chain polypeptide as a
secretory component.
 It binds specifically to poly Ig
receptor (Fc region binds)
 It is an integral membrane glycoprotein

3. IgE
 Mediates any immediate hypersensitivity reactions
 In regard to fever, asthma, hives and anaphylactic shock.
 Binds to fc receptors on membranes of basophils and tissue mast cells.
  Cross-linkage between antigen and receptor bound antibody induces degranulation of
basophils and mast cells
 Necessary for anti-parasitic defense.
4. IgD
 Constitutes 0.2% of total serum Ab proteins
 Together with IgM, they are membrane bound complexes expressed by mature B cells
 To function in activation of B cells by antigen
 No biological effector function identified yet.
5. IgG

 Consists of 80% of total serum Ab proteins

 There are four types: IgG1, IgG2, IgG3 and IgG4
 Half-life is 7 days
 It is the only antibody which can pass through the placenta and enable transfer of
immunity from mother to foetus.

Antigenic determinants of
antibodies

1. Isotype
 Determined by the constant region sequence of the heavy chain
 The five human isotypes; IgM, IgG, IgA, IgD and IgE have structural and functional differences
 Constant region determinants defining each heavy chain class and sub-class within a species
 Studying a class of antibodies with the extent of its similarity
 All members would carry the same constant region of the chain
 2. Allotype
 Based on genetic differences
between individuals
 These are antibodies
belonging to individuals of the same species but having certain genetic differences
 Changes in minor amino acids
 Either the heavy chain or light chain can be present or absent in any individual of a species
 Variation in constant heavy chain and light chain
 The sum of all individual allotypic determinants results in an allotype

3. Idiotype
 Generated by conformation of the heavy and light chains
 There is a unique amino acid sequence of variable domains in the given antibody
 It can function not only as an antigen binding site but also as a set of antigenic determinants
 These are different immunoglobulins arising from the same individual
 Each individual antigenic determinant of the variable region is termed as an idiotype

Isoantibodies
 1. ABO Blood Group
 Plasma membrane of RBC’s contain many antigens bound to its surface
 ABO antigens are a complex oligosaccharides present as glycoproteins and glycolipids
 The H molecule acts as a precursor coded by the H gene
 In A antigen – H contains N-acetylgalactosamine attached by enzyme GalNAc transferase
 In B antigen – H contains galactose attached by enzyme Gal transferase
 AB group contains both the products whereas O group contains none of the products and only H
antigen
 The hierarchy of dominance relationship is symbolized as
 Anti A antibody reacts with antigen A and Anti B antibody reacts with antigen B.
 Both antibodies react with type O blood group and none react with AB blood group
 AB group are universal recipients and O group is a universal donor

2. Rh Blood Group
 This is the Rhesus antigen which was first isolated in Rhesus monkeys
 Rh + and Rh – are the two types of antigens present on the surface of RBC’s

3. Erythroblastosis foetalis
 When a woman is pregnant, it’s possible that her baby’s blood type will be incompatible with
her own.
 This can cause a condition known as Erythroblastosis foetalis, where the mother’s white blood
cells (WBCs) attack the baby’s RBCs as they would any foreign invaders.
 Erythroblastosis foetalis is now known as hemolytic disease of the newborn.
 Babies who experience Erythroblastosis foetalis symptoms may appear swollen, pale,
or jaundiced after birth.
 There are two main causes of Erythroblastosis foetalis: Rh incompatibility and ABO
incompatibility.
 For example, if you’re type A and Rh positive, you have A antigens and Rh factor antigens on the
surface of your RBCs. Antigens are substances that trigger an immune response in your body.
 If you have AB negative blood, then you have both A and B antigens without the Rh factor
antigen.
 Rh incompatibility occurs when a Rh-negative mother is impregnated by a Rh-positive father.
The result can be a Rh-positive baby.
 Diagnosed by routine blood tests
 They may be given intrauterine blood transfusions to reduce anemia. 
 A preventive treatment known as RhoGAM, or Rh immunoglobulin, can reduce a mother’s
reaction to their baby’s Rh-positive blood cells. This is administered as a shot at around the 28th
week of pregnancy.
 The shot is administered again at least 72 hours after birth if the baby is Rh positive. This
prevents adverse reactions for the mother if any of the baby’s placenta remains in the womb.