CHAPTER 3

ANTIGEN & ANTIBODY

 Learning Outcomes
• Identify the characteristics of antigen.
• Describe immunogenic, weak immunogenic and not
immunogenic.
• Differentiate B cells and T cells in antigen
identification.
 What are ANTIGENS (Ags)?
• Term used to describe a substance that is specifically bound
by an antigen-recognizing molecule.
• Abs can recognize almost every antigen (metabolites,
sugars, lipids, carbohydrates, phospholipids, nucleic acids and
proteins).
• In contrast, T cells only recognize peptide antigens.
• Although Ags are large molecules, Abs and TcR can only
recognize specific regions called the antigenic
determinant (aka epitope).
• In an antibody-antigen reaction the epitope complements
the paratope perfectly.
 Hapten
• Haptens are small molecules which induce an immune
response when coupled to a carrier molecule (foreign
protein).
• e.g.: dinitrophenyl, aminobenzene, sulphonate,
arsonate.

• Hapten-carrier molecule, unlike free hapten, can acts

as an immunogen  substances that generate
immune response.
Haptens and Hapten-carrier
Conjugate Hapten-carrier
molecule
 Immunogenicity of Haptens

• Immunogenicity  Ability of a particular substance, such as an antigen or

epitope, to provoke an immune response in the body of a human or animal.
• Small molecule would not antigenic by itself.
• Becomes antigenic when bound to “carrier molecule”.
 Antigenicity and
Immunogenicity
• Antigenicity  The strength in inducing an immune
response.
• Immunogens are always antigens BUT antigens are not
necessarily immunogenic.
• Characteristics of antigen to become immunogen:
Foreign substance (organic and non-organic)
Chemically complex (heterogeneity in the building blocks;
proteins vs polysaccharides)
High molecular weight (10 kDa above are usually highly
immunogenic)
Adjuvant  Substances that enhance immune response
to Ag
 Chemical Nature of
Immunogens
• Proteins
• Polysaccharides
• Nucleic acids
• Lipids
 Antigen-recognizing Molecules
• 3 classes of molecules recognize foreign
antigens; antibodies, the T cell receptor (TcR) and
major histocompatibility complex (MHC) molecules.
• Antibodies act as membrane-bound receptors in the
recognition phase of an immune response.
• They can also be produced in secreted form by
plasma cells, bind antigen and trigger effector
responses that eliminate the antigens.

Major
histocompatibility
complex (MHC) on
APC
 ANTIBODIES
(IMMUNOGLOBULINS)
• Also known as immunoglobulins.
• Is a globular glycoproteins.
• The heavy and light chains are polypeptides.
• The chains are held together by disulphide
bridges.
• Each Ab has 2 identical Ag binding sites
 variable regions.
• The order of amino acids in the variable
region determines the shape of the binding
site.
• Ab binds epitopes on a wide range of intact
molecules; protein, carbohydrate, lipid.
• Antibodies act as
membrane-bound
receptors on the surface of
B cell in the recognition
phase of an immune
response.
• They can also be produced
in secreted form by plasma
cells, bind antigen and
trigger effector responses
that eliminate the
antigens.
 Ab: The Basic Recognition
Unit

Paratope

Antigen Marker molecule Antibody

Page  12
 Immunoglobulins
Isotypes/Classes
• Classes (isotype) and
subclasses (subtype) are
based on differences in the
amino acid sequences in
heavy chain C domains.
• Heavy chains are
designated by greek letters
(α, μ, , δ, ε, γ).
• Five classes isotypes; IgG,
IgA, IgM, IgE and IgD.
 Number of
Type Ag binding Site of action Functions
sites
• Increase macrophage activity
• Blood • Antitoxins
IgG 2 • Tissue fluid • Agglutination
• Can cross placenta • Coat microbes, speeding their uptake by other cells in the
immune system.

• Blood • Agglutination
IgM 10
• Tissue fluid • Very effective at killing bacteria.

Secretions (saliva, tears, • Guarding the entrances to the body.

IgA 2 or 4 small intestine, vaginal, • Stop bacteria adhering to host cells
prostate, nasal, breast milk) • Prevents bacteria forming colonies on mucous membranes

• Protect against parasitic infections

IgE 2 Tissues • Activate mast cells  HISTAMINE
• Causes allergy
• Remains attached to B cells and plays a key role in initiating
IgD 2 Part of B cell receptor early B cell response.
• Activates basophil and mast cells
Antigenic Determinants
(Epitope)
• Epitope  The part of Ag (at it’s
surface) that recognized by Ab.
• Epitopes can either be due to
covalent structure (forming a linear
determinant) or tertiary structure
through juxtaposition of amino
acid residues (forming a
conformational determinant).
• The paratope can occupy a linear
determinant of up to 6 amino
acids.
• Some linear determinants are
hidden in the native folded protein
and only bind when the protein is
denatured (externalized).
• Alterations to covalent structure
through phosphorylation or
proteolysis can produce new
epitopes named neoantigenic
determinants.
Antigenic Determinant
 Antibody-Antigen
Interactions
• Binding of antibody to antigen is dependent on
 hydrogen bonds
 lectrostatic attractions
 Van der Waals forces
 hydrophobic interactions

• These bonds are weak compared to covalent bonds

but the large number of weak bonds result in a stable
complex.

• Antibody-antigen binding is reversible.

 General functions of Ab
• Block toxins, e.g. those causing diphtheria and
tetanus.
• Neutralizing antigen.
• Immobilizing bacteria by attaching to bacterial
flagella making them less active and easier for
phagocytes to engulf.
• Lead to agglutination of bacteria making them less
likely to spread.
• Activating complement.
• Enhancing phagocytosis.
Effect of Antibodies
 MAJOR HISTOCOMPATIBILITY
COMPLEX (MHC)
• MHC  group of genes involved in the immunological
recognition of self and non-self in animal species (Penn
and Potts, 1999).
• The gene for MHC is located in chromosome 6.
• The name of the MHC varies with the species, such that
in humans the MHC is termed HLA (Human Leukocyte
Antigen) and in the mouse it is called H-2.
• MHC genes are very polygenic (contains several different
genes) and polymorphic (poly = many; morphic = shape,
structure).
• Plays an important role during transplant rejection.
 Classes of MHC
1) MHC Class I
• Glycoproteins expressed on all nucleated
cells
• Major functions to present processed
antigens to cytotoxic T cells
2) MHC Class II
• Glycoproteins expressed on antigen
presenting cells (APC)
• Major function to present processed
antigens to helper T cells
MHC Class I and II
 Antigen Presenting Cells
(APCs)
•Cells with the capacity to capture, process and
present antigenic peptides to helper T cells.
•APCs deliver co-stimulatory signal to T cells
leading to proper activation.
•Only APCs can activate a naive T cell
 Dendritic cells, macrophages, B cells.
Spleen
& LN
Pathway of Ag Presentation
 T CELL RECEPTOR (TCR)
•T cell receptor  surface
molecule analogous to
part of Ab.
•Diversity is generated by
rearrangement of TCR
gene locus.
• Recognizes and binds
mainly on peptide
antigens, derived from the
pathogen’s proteins.
 T CELLS RECEPTOR (TCR)
•T cell receptors (TCR) is antigen specific.
•CD4 (Th cells) or CD8 (Tc cells)

CD4 CD8
TCR TCR

CD4 T cell/ CD8 T cell/

helper T cell cytotoxic T cell
 TCR: CD4
•T cells that express CD4+ molecule on their
surface  Helper T cells.
•Recognition of exogenous antigen.
 bacteria
 extracellular Ag
•Recognize MHC class II molecule present on
APC.
•Modulate cellular immunity by secreting
cytokines that modulate:
 B cell activation
 Ab secretion
 Macrophage and dendritic cell
activation
 Cellular chemotaxis and inflammation
 TCR: CD8
•T cells that express CD8+ molecule on their
surface.
•30% of T cells in the periphery.
•Destroy cells infected by intracellular pathogens
and cancer cells.
•Recognize class I MHC molecules (nucleated body
cells) expose foreign proteins.
•Tc cells release perforin and granzymes;
proteins that form pores in the target cell
membrane and causing cell lysis and/or apoptosis.
CD8 in ACTION!!!
HELLO.....QUIZ~~
1. List THREE antigen
recognizing molecules.
2. List THREE antigen
presenting cells.
3. MHC Class I is expressed
on __________.
4. MHC Class II is expressed
on ________.
5. List all classes of
immunoglobulin.
6. State ONE function of
antibody.
7. Helper T cell also known as
_______.
8. Cytotoxic T cell also known
as _______.
9.Fill in the blank with suitable
answers.
MHC Class I Characteristics MHC Class II
Present
Nature of ag
? (a) exogenous
presentation
antigen
Cytotoxic T cell Responsive T cell ? (b)

? (c) End result ? (d)