Function and structure of Antibodies and MHC

Dr. Muhammad Zubair

FCPS chemical pathologist
 Definitions
Genome.
The total genetic material contained within the cell.
Gene.
(Gr, Gennan to produced) the smallest biologic unit of
heredity material.
Locus.
Specific site where a gene is located on a chromosome.
Allele.
Genes located at corresponding loci on a pair of
chromosomes.
 Definitions
Region.
The term region is used to designate a
chromosome segment occupied by different loci.
Polymorphic.
The genes that code for multiple alleles at a
given locus.
 Definitions
Dominant Gene.
The gene that expresses whether homozygous or
heterozygous on pair of chromosome.
Codominant Genes.
When both the genes in heterozygous state are
expressed on pair of chromosome. e.g. Blood Group AB.
Recessive Genes.
A gene that expresses when it is present in homozygous
state or when it is transmitted by both parents.
 Definitions
Histocompatibilty Genes.
A gene that determines the specificity of tissue
antigenicity and thus the compatibility of donor and
recipient in tissue transplantation and blood transfusion.
Haplotype.
A set of genetic determinants located on a single
chromosome.
H.L.A.
Human Leucocyte antigen.
 MAJOR HISTOCOMPATIBILTY COMPLEX

The genes complex present on small segment of

chromosome No.6. is termed as major
histocompatibility complex.
CLASSIFICATION OF MAJOR HISTOCOMPATIBILTY COMPLEX

Three types of antigen are present.

 Class I antigen.
 Class II antigen.
 Class III antigen.
 GENETICS OF MHC CLASS I
ANTIGEN
• These are encoded by A,B,C loci.
• These are written as HLA-A,HLA-B,HLA-C (HLA
is followed by the name of locus).
• After the name of the locus, gene number is
written e.g. HLA- A2.
• If the gene is still being under evaluation then
w is written before the number e.g. HLA-
BW4, (w stand for international workshop).
GENETICS OF MHC CLASS I ANTIGEN

•  MHC class I molecules are present on the

surface of all nucleated cells and platelets.
• These are not present on red blood cells.
• Present the peptide antigen to CD8+ T cells
 GENETICS OF CLASS II ANTIGEN

• These are encoded by D locus.

• D locus is further sub divided in to DR,DP,DQ.
• Nomenclature is like class I antigen e.g. HLA-
DR6,HLA-DQ4.
 GENETICS OF CLASS II ANTIGEN

• In addition MHC region also contain non

classical genes such as DM, DO, LMP and TAP
that help in antigen processing and
presentation.
• MHC-II proteins are located on the surface of
APC. (Antigen Presenting Cells)
• Present the peptide to CD4+ T cells
 GENETICS OF CLASS III ANTIGEN

• Class III are the products of complement

component.
• Comprises of genes that code for
complement factors, heat shock proteins
(HSP), tumor necrosis factor (TNF α and β),
steroid 21 hydroxylases etc
 INHERITANCE OF MHC(HLA)
• The person inherits two genes from each
locus.
• Maximum a person can have 6 genes from
class I loci and 6 genes from class II loci.
• Minimum a person can have 1 gene from each
locus of class I and Class II antigen (inbred
animals, uniovular twin, repeated intra family
marriages).
• These genes are polymorphic therefore
produced distinct antigens.
INHERITANCE MHC
STRUCTURE OF CLASS I ANTIGENS

• These consist off polymorphic α and non

polymorphic β2 microglobulin.
• Extra cellular portion of class I antigen
consist of α1, α2, α3 and β2 microglobulin
domains.
• α1 and α2 forms the cleft for antigen
binding.
STRUCTURE OF CLASS I ANTIGEN
 STRUCTURE OF CLASS II ANTIGEN
• It is a heterodimer consisting of noncovalently
associated α chain and β chain, both chains
are polymorphic.
• Extra cellular portions of the α and β chains
have two domains each: α1, α2 and β1, β2.
• Class II molecules present exogenous antigen
e.g. extra cellular microbes, soluble proteins.
• These antigen are processed and are
presented on surfaces of the cell along with
Class II proteins.
MHC CLASS II ANTIGEN
 MHC CLASS II ANTIGEN

• Class II are only present on antigen

presenting cells (APC’s) & B Cells.
• CD4 cells can recognized antigen with the
help of Class II antigen.
• Class II antigen expression can be
induced on other cells like endothelial
cells and fibroblasts by the IFN-Ý.
 Regulation of MHC expression
• Transcription factors.MHC genes have
promoter sequence at their 5’ end which are
regulated by certain transcription factors such
as CIITA(Class II transactivator) and RFX (both
bind to MHC II molecules and increase their
transcription).
• Defect in CIITA and RFX- leads to Bare
lymphocyte syndrome.
 Cytokines:
• IFN-γ activates both MHC-I and II promoter
genes.
• IL-4 increases the expression of MHC II
molecules in resting B cells
• Corticosteroids and prostaglandins decrease
the expression of MHC II molecules.
• Viral infection: Some viral antigens inhibit
various components of MHC I (e.g. adenovirus
protein inhibit TAP, cytomegalovirus protein
inhibit β2 microglobulin). As a result, MHC-I
expression is suppressed.
 HLA Nomenclature
• There are a number of ways of writing an HLA antigen.
For example, it may be expressed as HLA-DR3, HLA-DRI7,
HLA-DRB* 03 or HLA- DRBI*0301. These could all refer to
the same antigen.
• Firstly, “HLA” is the name for the gene cluster which
tends to be inherited en-bloc.
• Second part -e.g. DR- is the name of the specific locus.
There are 6 loci normally referred to. These are A, B, C,
DR, DQ and DP
• Third part, the number, e.g. 3, 17, 03, 0301, refers to the
actual antigen at the locus.
 HLA TYPING
• In this test, donor’s antigens expressed on the surface
of leukocytes or their genes are matched with that of
the recipient.
• The closer the HLA antigens on the transplanted organ
match the recipient, the more likely that the recipient’s
body will not reject the transplant.
• Value of HLA matching between donor and recipient
varies in different solid organ transplantation.
• In kidney transplants, there is substantial benefit if all
the polymorphic HLA alleles are matched.
• Every person inherits each of the following
antigens from each parent:
• HLA-A antigen
• HLA-B antigen
• HLA-C antigen
• HLA-DR antigen
• HLA-DQ antigen and
• HLA-DP antigen 
• When performing an HLA typing test for a
kidney transplant, the following HLA antigens
are looked at:
• HLA-A
• HLA-B
• HLA-DR
• Six HLA antigens are looked at for each person
• By analyzing which six of these HLA-antigens
both the donor and recipient have, scientists
are able to determine the closeness of tissue
matching.
• A six-antigen match is the best compatibility
between a donor and recipient.
• This match occurs 25% of the time between
siblings who have the same mother and father
 METHODS OF HLA TYPING

• Serological technique, tissue

typing.
• Cellular techniques.
• DNA typing method.
• DNA analysis by PCR.
 SIGNIFICANCE OF HLA
• Inflammatory diseases, including ankylosing
spondylitis and several post infectious
arthropathies, all associated with HLA-B27.
• Inherited errors of metabolism, such as 21-
hydroxylase deficiency (HLA-BW47) and
hereditary hemochromatosis (HLA-A).
• Autoimmune disease, including autoimmune
endocrinopathies, associated mainly with alleles
at the DR locus.
• HLA antigen are Major Transplant Antigen. i.e.
success of transplant depends upon them.
 SIGNIFICANCE OF HLA
• Class I antigen are responsible for immune
recognition.
• Class I antigen mediate elimination of virally infected
cells.
• Class II antigen are concerned with immune response
at many steps. e.g. antigen presentation to CD4 cells.
• Class II antigen are play major role in graft versus
host disease.
• HLA antigen are used to decided disputed paternity.
• These antigens play a major role in immune
tolerance.
• These antigens are also concerned with body defense
against tumors.
• A number of diseases have been found to
occur at a higher frequency in individuals with
certain MHC haplotypes. Most prominent
among these are ankylosing spondylitis (B27),
celiac disease (DR3), Reiter's syndrome (B27)
 Disease associations with Class I HLA

• Ankylosing spondylitis (B27)

• Reiter's disease (B27),
• Acute anterior Uvietis (B27)
• Psoriasis vulgaris (Cw6)
 Disease associations with Class II HLA

• Hashimoto's disease (DR5), Primary myxedema

(DR3), Graves thyrotoxicosis (DR3), Insulin-
dependent diabetes (DQ2/8), Addison's
disease (adrenal) (DR3),Goodpasture's
syndrome (DR2), Rheumatoid arthritis (DR4),
Juvenile rheumatoid arthritis (DR8), Sjogren's
syndrome (DR3), Chronic active hepatitis
(DR3), Multiple sclerosis (DR2, DR6), Celiac
disease (DR3), Dermatitis herpetiformis (DR3
IMMUNOGLOBULINS/
ANTIBODIES
 ANTIBODY
• These are glycoprotein's produced in
response to an antigen and have property of
combining with the antigen in some
detectable way.
• These are also termed as immunoglobulins.
 Structure of Antibody
Antibodies can be made against proteins,
carbohydrates, lipids and nucleic acids.

 Antibodies to nucleic acids and lipids can be

found in autoimmune diseases.
 Antibodies to small organic
molecules can cause allergies to drugs.
 Structure of Antibody
• Antibodies are heavy (~150 kDa) globular plasma
proteins.
• The basic structure of all antibodies are same
• There are four polypeptide chains: two identical
heavy chains and two identical light chains
connected by disulfide bonds. 
• Light Chain (L) consists polypeptides of about
22,000 Da and Heavy Chain (H) consists larger
polypeptides of around 50,000 Da or more. 
• There are five types of Ig heavy chain (in
mammal) denoted by the Greek letters: α, δ,
ε, γ, and μ
• There are two types of Ig light chain (in
mammal), which are called lambda (λ) and
kappa (κ).
Structure of Antibody
• An antibody is made up of a variable region
and a constant region, and the region that
changes to various structures depending on
differences in antigens is called the variable
region, and the region that has a constant
structure is called the constant region.
 CLASSIFICATION OF
IMMUNOGLOBULINS/ANTIBODIES

• Depending upon type of heavy chain

• IgM. Contains µ (Meu) heavy chain
• IgG. Contains Gamma heavy Chain
• IgA. Contains α alpha heavy Chain
• IgD. Contains Delta heavy Chain
• IgE. Contains € Epsilonheavy Chain
 IMMUNOGLOBULINS IgM
• Macroglobulin having M.wt.970,000.
• Exists in monomeric & Pentmeric form.
• In pentmeric form 5 molecules are joined by
invariant J Chain.
• Five antigen binding sites instead of ten due to
steric hindrance & St.rigidity.
• Cannot cross placenta.
• Efficient agglutinating / complement fixing.
• An early antibody produced in immune
response.
 IMMUNOGLOBULINS IgA
• Accounts for 10-15% of serum Ig.
• Principal antibody class conferring mucosal
protection, secreted in body secretions.
• Produce by B lymphocytes in sub mucosal lymphoid
tissues.
• It has two subclasses IgA1 & IgA2.
• Exists in monomer, dimer and trimer form.
• Secreted as dimer linked by secretory component
produced by mucosal cells, which give protection
from proteolysis.
• Higher levels are seen in chronic liver disease and IgA
nephropathy (Berger’s disease).
 IMMUNOGLOBULINS IgG

• Major immunoglobulin 70-75% of serum.

• Molecular weight (146,000 -150,000).
• Distributed evenly between vascular & extra
vascular pool.
• Major antibody of secondary response.
• It can cross placenta.
• Is subdivided into four subclasses. IgG1-IgG2-IgG3-
IgG4
 IMMUNOGLOBULINS IgD
• Accounts for <10% serum Ig.
• Present on B lymphocytes which also
bear IgM.
• Precise biological function is not known.
• Probably has some role in insulin
resistance/class switching.
 IMMUNOGLOBULINS IgE
• Present in trace amount in serum (0.004%),
and in monomer form.
• Important Ig of allergic / hypersensitivity
reactions.
• Basophil / mast cells bear high affinity
receptors specific for Fc portion.
• Bound Ig combined with antigen results in
degranulation of basophil / mast cells.
• High level are seen in parasitic infection.
 Properties of Immunoglobulins
lgG lgA lgM lgD lgE
Sedimentation coefficient (s) 6-7 7 19 7- 8 8
Molecular weight (approximate) 150,000 160,000 900,000 180,000 190,000
or
400,000
Complement fixation (Classic) + 0 ++++ 0 0

Serum Concentration 1000 200 120 3 0.05

(Approximate; mg/dL)
Serum half-life (days) 23 6 5 3 2
Placental transfer + 0 0 0 0
Mast cell or basophil ? 0 0 0 ++++
degranulation
Bacterial lysis + + +++ ? ?
Antiviral activity + +++ + ? ?
 Function of the Immunoglobulins

• Neutralisation.
• Agglutination.
• Complement.
• Opsonisation.
• Placental transfer.
 IMMUNOGLOBULINS IgG
Increases in:
–Chronic granulomatous infections
– Infections of all types
– Liver disease
– Malnutrition (severe)
– Dysproteinemia
– Disease associated with hypersensitivity granulomas,
dermatologic disorders, and IgG myeloma
 IMMUNOGLOBULINS IgG
Decreases in:

a)Agammaglobulinemia
b)Lymphoid aplasia
c)Selective IgG, IgA deficiency
d)Bence Jones proteinemia
e)Chronic lymphoblastic leukemia
 IMMUNOGLOBULINS IgA
Increases in:
a) Waldenström's macroglobulinemia
b) Trypanosomiasis
c) Actinomycosis
d) Malaria
e) Infectious mononucleosis
f) Lupus erythematosus
g) Rheumatoid arthritis
h) Dysgammaglobulinemia (certain cases)
 IMMUNOGLOBULINS IgA
Decreases in:

 a) Agammaglobulinemia
b) Lymphoproliferative disorders (certain cases)
c) Lymphoid aplasia
d) Dysgammaglobulinemia
e)Chronic lymphoblastic leukemia
 IMMUNOGLOBULINS IgE

Increases in:
 a) Atopic skin diseases such as eczema
b) Hay fever
c) Asthma

d) Anaphylactic shock
e) IgE-myeloma
Decreases in:
 a) Congenital agammaglobulinemia
 b) Hypogammaglobulinemia due to faulty metabolism
or synthesis of immunoglobulins
 IMMUNOGLOBULINS IgD
• The function of IgD is unclear. A high level may
mean IgD multiple myeloma, which is much
less common than IgA or IgG multiple
myeloma.
 IMMUNOGLOBULINS IgM
• Low levels occur in multiple myeloma, some
types of leukemia, and in some inherited types
of immune diseases.
• Some patients are asymptomatic, while others
develop serious infections.
• This deficiency is usually discovered during the
investigation of other conditions.
 IMMUNOGLOBULINS IgM

• HIgh level indicate that IgM MGUS,

Waldenstrom's macroglobulinemia, early viral
hepatitis, mononucleosis, rheumatoid
arthritis, kidney damage (Nephrotic
syndrome), or a parasite infection is present.
• IgM antibodies are the type that form when
an infection occurs for the first time, high
levels of IgM can mean a new infection is
present.
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