IMMUNOLOGI

Antibody structure and development

Putu Yustiantara
 Antibodi
• Antigen specific proteins produced by plasma
cells
• Belong to immunoglobulin superfamily
• Located in blood and extravascular tissues,
secretions and excretions
• Bind pathogenic microorganism and their
toxins in extracellular compartments
• Secreted form of immunoglobulins
 Imunoglobulin
• Antigen specific proteins produced by B
lymphocytes
• Belong to immunoglobulin superfamily
• Bound to surface of B lymphocytes
– Function as binding (receptor) sites for
specific antigens
• Antigen receptor sites on mature B
lymphocytes
– IgM
– IgD
 CLASSES (ISOTYPES) OF
IMMUNOGLOBULINS

• Classes based on constant region of heavy chains

– Immunoglobulin A (IgA)
– Immunoglobulin D (IgD)
– Immunoglobulin E (IgE)
– Immunoglobulin G (IgG)
– Immunoglobulin M (IgM)
• Differentiation of heavy chains
– Length of C region, location of disulfide bonds, hinge region,
distribution of carbohydrate
• Classes have different effector functions
 STRUCTURE OF ANTIBODIES
• Antibodies are glycoproteins composed of
– Polypeptide chains and carbohydrate

• Monomeric structure
– Polypeptide chains
• 2 identical heavy chains
• 2 identical light chains
– Polypeptide chains joined by disulfide bonds
– Carbohydrate
 STRUCTURE OF ANTIBODIES
• Polypeptide chains have variable and constant regions
– Variable
• N (amino)-terminal of polypeptide chain
• Antigen binding site
– Constant
• C (carboxyl)-terminal of polypeptide chain
• Binding sites for cell surface receptors and
complement
• Structure represented by the letter “Y”
• Y shaped molecule cleaved by protease papain
– Fragment antigen binding (Fab)
– Fragment crystallizable (Fc)
 ANTIGEN BINDING SITES OF
IMMUNOGLOBULINS

• Antigen binding sites formed from hypervariable regions

– Heavy chain V domain
– Light chain V domain

• Hypervariable regions of V domains

– Amino acid sequence differences concentrated
– Flanked by less variable framework regions
– Three hypervariable regions in each V domain
– Hypervariable regions also called
• Complementarity-determining regions (CDR)
 ANTIGEN BINDING SITES OF
IMMUNOGLOBULINS

• Antigen binding sites vary with size and shape of antigen

• Part of antigen to which antibody binds

– Antigenic determinant (Epitope)

• Antigen-Antibody binding based on non-covalent forces

– Hydrogen bonds

• Affinity
– Strength of binding of one molecule to another by a single binding site

• Avidity
– Overall strength of binding between two molecules
 IMMUNOGLOBULIN DIVERSITY IN
B-CELLS BEFORE ENCOUNTER WITH
ANTIGEN
• Immune system capable of producing a limitless
number of different immunoglobulins/antibodies
• Mechanism
– Genes for IG organized differently
• Genes exist as nonfunctional segments
– Variable (V), Joining (J), Diversity (D), Constant
(C)
• Genes are inherited in this form
– Germline form (germline configuration)
Antibody Diversity
B lymphocyte development
 IMMUNOGLOBULIN DIVERSITY
IN B-CELLS BEFORE ENCOUNTER
WITH ANTIGEN
• Expression
– Gene segments must be rearranged into functional
gene
• Gene Rearrangement
– Takes place during development of B-cells
– Mechanism of somatic recombination
• Genes for IG located at 3 chromosomal locations
– Heavy chain locus on chromosome 14
– Kappa light chain locus on chromosome 2
– Lambda light chain locus on chromosome 22
CONSTRUCTION OF LIGHT CHAIN
AND HEAVY CHAIN VARIABLE
REGIONS
• Light chain
– Constructed from 2 segments
• 1 (V) segment
• 1 (J) segment

• Heavy chain
– Constructed from 3 segments
• 1 (V) segment
• 1 (D) segment
• 1 (J) segment
 MECHANISMS OF GENETIC
DIVERSITY IN V-REGION OF
IMMUNOGLOBULINS
• Random combination of
– V and J segments in light chain genes
– V, D and J segments in heavy chain genes

• Addition of P (palindromic) and N (non-templated)

nucleotides at junctions of gene segments during
recombination
– Junctional diversity

• Association of H and L chains in different combinations

Diversity calculations
THE PRIMARY HUMORAL IMMUNE
RESPONSE
• Immune response initially produces IgM antibodies then switches to
IgG antibodies

• Question
– Why switch from IgM to IgG?

• Answer
– Limited effector mechanisms for IgM
– Range of effector mechanisms for IgG

• Mechanism
– Isotype or class switching
B lymphocyte development (2)
 ISOTYPE OR CLASS SWITCHING

• Process by which B cell changes class of IG produced while

preserving antigenic specificity

• Involves somatic recombination which attaches different

heavy chain constant region to variable region

• Occurs only during active immune response

• Mechanisms involves recombination between

– Switch sequences (regions)
Class switching among constant regions: generation
of IgG, IgA and IgE with same antigenic determinants
—idiotypes
 Regulation of Ig gene transcription
Each lymphocyte rearranged gene has regulatory sequences that
control gene expression
• Promoters: initiation sites of RNA transcription
• Enhancers: upstream of downstream that transcription from the
promoter sequence
• Silencers: down-regulate transcription in germline cells
• Gene rearrangement brings enhancer and promoter regions close
together and eliminates silencer regions allowing transcription
 CLASSES, SUBCLASSES AND
PHYSICAL PROPERTIES OF
IMMUNOGLOBULINS
Classes Subclasses
IgG IgG1, IgG2, IgG3, IgG4

IgA IgA1, IgA2

IgM

IgD

IgE

Subclasses are numbered according to plasma

concentration
FUNCTIONS AND PROPERTIES OF
ANTIBODY

• Neutralization
– Direct inactivation of pathogen or toxin thereby
preventing its interaction with human cells

• Opsonization
– Coating of pathogens for more efficient phagocytosis

• Activation of complement
– More efficient phagocytosis
– Direct killing
IgM ANTIBODY OF THE IMMUNE
RESPONSE

• First isotype produced in primary response

– May or may not be produced in secondary response
– Produced before B cells undergo somatic
hypermutation
• Occurs as pentamer with J chain
– Found primarily in blood and lymph
• Multiple binding sites confers high avidity and
compensates for low affinity of monomers
• Highly effective in complement activation
• Functions as rheumatoid factor
 IgG ANTIBODY OF THE IMMUNE
RESPONSE

• Second isotype produced in primary response

• Primary isotype of
– Secondary immune response
– Memory immune response
• Represents approximately 75% of total serum IG
• Four subclassses (1-4)
– Different effector functions
• Transported across placenta
• Functions as rheumatoid factor
 IgA ANTIBODY OF THE IMMUNE
RESPONSE
• Two subclasses (IgA1 and IgA2) and two forms (monomeric and
dimeric)

• Monomeric
– Located in blood and extracellular spaces
– Predominately IgA1
• Ratio of IgA1 to IgA2 is 10:1
– Functions as rheumatoid factor

• Dimeric
– Located in mucous membranes and secretions
– Predominately IgA2
– Ratio of IgA2 to IgA1 is 3:2
– J chain like IgM
IgE AND IgD ANTIBODIES OF THE
IMMUNE RESPONSE

• IgE
– Binds with high affinity to receptors on mast cells,
basophils and activated eosinophils
– Longer half-life when cell bound
– Initiates a strong inflammatory reaction to parasites
– Involved in allergic reactions

• IgD
– Antigen receptor on mature B-cells
– No other known function
terimakasih