IMMUNOLOGY

POLYMORPHISM OF MHC GENES,

ROLE OF MHC ANTIGEN IN IMMUNE RESPONSE,
MHC ANTIGENS IN TRANSPLANTATION.

PRESENTED BY:
MONISHA R
22BC13
II-Msc BIOCHEMISTRY
MAJOR HISTOCOMPATIBILITY COMPLEX

 MHC molecules were initially discovered during studies

aimed at understanding the molecules responsible for
rejection of transplanted tissues.
 The term MHC actually refers to a region of the genome
that encodes a number of genes that play an important
role in tissue transplantation.
 MHC was discovered by Peter Gorer and George Sneell
(1940)
 MHC GENES
 Antibodies or B-cell receptors, which can recognize an antigen
alone, T-cell receptors only recognize pieces of antigen that are
positioned on the surface of other cells. These antigen pieces
are held within the binding groove of a cell surface protein
called the major histocompatibility complex (MHC)
molecule.
 The MHC genes in the region encode proteins that determine
whether a tissue transplanted between two individuals will be
accepted or rejected.
 There are three main classes of MHC molecules:
 MHC class I
 MHC class II
 MHC class III.
 MHC GENES
 Class I MHC genes encode glycoproteins expressed on the
surface of nearly all nucleated cells; the major function of the
class I gene products is presentation of endogenous peptide
antigens to CD8 T cells.
 Class II MHC genes encode glycoproteins expressed
predominantly on APCs (macrophages, dendritic cells, and B
cells), where they primarily present exogenous antigenic
peptides to CD4 T cells.
 Class III MHC genes encode several different proteins, some
with immune functions, including components of the
complement system and molecules involved in inflammation.
 MHC-I MOLECULE
 MHC-I molecule is a Heterodimers.
 It has two noncovalently bound chains:
Alpha chain:
- encoded in the MHC transmembrane
- 3 domains(1, 2, 3)
β2-microglobulin:
- not encoded in MHC
- not transmembrane
- 1 domain
 Class I MHC cannot be expressed without β2-microglobulin
 α1 and α2 domains form the peptide binding cleft.
 3 domain performs a structural role but has no direct role in
peptide binding.
 CD8 binds to the α3 domain.
 MHC-II MOLECULE
 Heterodimers
 It has two noncovalently bound chains
Alpha and Beta chains:
- both encoded in the MHC
- both transmembrane
- Both chains are necessary for expression of class II
MHC
 α1 and β1 domains from the peptide-binding cleft
 α2 and β2 domains perform a structural role in
peptide binding.
 The β2 domain is bound by CD4
 ORGANIZATION OF MHC
COMPLEX IN MOUSE AND HUMAN
 In humans MHC genes present in
the chromosome 6.  In mouse MHC genes present in the
 In humans MHC genes called chromosome 17.
Human HLA complex.  In mouse MHC genes are called
 Class I= A,B and C(also called H2-complex.
HLA-A,HLA-B and HLA-C)  Class I=K, D and L molecules(also
Ag (peptide) presentation to CD8+ called H-2K, H-2D and H-2L.
cells  Class II= A and E(also called I-A
 Class II=DP,DQ and DR(also called and I-E)
HLA-DP,HLA-DQ and HLA-DR)  Class III= Complement proteins,
Ag(peptide) presentation to CD4+ tumor necrosis factor(TNFs)-α, β
cells
 Class III= Complement proteins,
 POLYMORPHISM OF MHC
GENES
 The MHC is highly polymorphic, meaning that there are many different
alleles (variants) of each MHC gene. This polymorphism is thought to be
beneficial for the immune system because it allows it to recognize a wide
variety of foreign antigens.
 The polymorphism of MHC genes is thought to be caused by a number of
factors, including:
• Gene duplication: The MHC genes are thought to have duplicated many
times over the course of evolution.
• Mutation: Mutations in MHC genes can lead to changes in the amino acid
sequence of the encoded proteins. These changes can affect the ability
of the proteins to bind to antigens.
• Recombination: MHC genes are often recombined during meiosis, the
process by which gametes (sex cells) are formed. This recombination can
 MHC POLYMORPHISM
 The loci that encodes class I and Class II MHC molecules are
the most polymorphic known in higher vertebrates.
 Within any species, there are many different alleles for each
class I and class II MHC molecule.
Human:
 HLA Class-I genes: A (240), B (470), C (110) alleles (1.2 x )
different class-I combinations
 HLA Class-II genes:
DP= DPB1 (96) alleles
DQ= DQA1 (22), DQB1 (49) alleles
DR= DRB1 (304), DRB1 (1), DRB1 (35), DRB1 (11), DRB1 (15)
alleles
1.8 x different Class II combinations, and
(1.2 x ) x (1.8 x ) = 2.25 x different combinations of Class I and
Class II possible combinations
 ROLE OF MHC ANTIGEN IN
IMMUNE RESPONSE
 Major histocompatibility complex (MHC) antigens are proteins that
are essential for the immune system to function properly. They are
found on the surface of all cells in the body and help the immune
system distinguish between self and non-self cells.
 When a foreign antigen, such as a bacteria or virus, enters the
body, it is captured by immune cells called dendritic cells.
 When a T cell receptor binds to an MHC molecule that is displaying
a foreign antigen, the T cell is activated and begins to produce
antibodies or other immune molecules that attack the foreign
antigen.
 Class I MHC antigens present antigens to cytotoxic T cells, which
are responsible for killing cells that are infected with viruses or
bacteria. Class II MHC antigens present antigens to helper T cells,
which help to activate other immune cells.
 ENDOGENOUS PATHWAY
 Endogenous pathway processed and presented the endogenous
antigens i.e. those generated within cell e.g. Viral infected cells,
tumour cells and intracellular pathogens (M. tuberculosis,
Histoplasma capsulatum).
 The processed antigen is presented on the cell membrane with
MHC-class I molecule which is recognized by CD8 + Tc-cell for
degradation.

 STEPS INVOLVED IN ENGOGENOUS PATHWAY:

 Proteolytic degradation of Ag (protein) into peptides.

 Transportation of peptides from cytosol to RER.
 Assembly of peptides with class I MHC molecules
 PROTEOLYTIC DEGRADATION OF PROTEINS INTO PEPTIDES:
• Intracellular proteineous antigen are larger in size to be bound to MHC molecule. So,
it is degraded into short peptides of about 8-10 amino acids.
• These proteins are degraded by cytosolic proteolytic system present in cell called
proteasome.
• Many proteins targeted for proteolysis have a small protein called ubiquitin attached
to them.
• Ubiquitin attached to them ubiquitin-protein complex consisting of 20S proteasome
and 19S regulatory component added to it.
• The resulting 26S proteasome cleaves peptide bonds which is ATP-dependent process.
• Degradation of ubiquitin protein complex is thought to occur within the central hollow
of the proteasome to release peptides.
 TRANSPORTATION OF PEPTIDES FROM CYTOSOL TO RER:
• Peptides generated in cytosol by proteasome are transported by TAP (transporter
associated with antigen processing) into RER (Rough endoplasmic reticulum) by a
process which require hydrolysis of ATP.
• TAP is consisting of two proteins, TAP1 and TAP2The optimal peptide length required
by class-I MHC for binding is nine, which is achieved by trimming the peptides with
the help of amino-peptidase present in RER. E.g. ERAP.
• TAP deficiency can lead to a disease syndrome that has both immune-deficiency and
auto-immunity aspects.
 ASSEMBLY OF PEPTIDES WITH CLASS I MHC MOLECULE:

• Within the rough endoplasmic reticulum (RER) membrane, a newly

synthesized class I chain associates with calnexin it is a molecular
chaperone, and ERp57 until β2microglobulin binds to the chain.
• The binding of β2-microglobulin releases calnexin and allows binding
to calreticulin and to tapasin, which is associated with the peptide
transporter TAP. This association promotes binding of an antigenic
peptide.
• Antigens in the ER can be further processed via exopeptidases such
as ERAP1, producing fragments ideally suited for binding to class I.
• Peptide association stabilizes the class I molecule-peptide complex,
allowing it to be transported from the RER to the plasma membrane.
 EXOGENOUS PATHWAY
 The endocytic pathway processed and present the exogenous Ag. i.e.
antigens generated outside the cells. E.g. Bacteria.
 At first APC phagocytosed, endocytosed or both, the antigen.
Macrophage and dendritic cells internalize the antigen by both the
process.
 While other APCs are non-phagocytic or poorly phagocytic. E.g. B cell
internalize the antigen by receptor mediated endocytosis.
 Then antigen is processed and presented on the cell surface along
with class-II MHC molecules which are recognized by CD4 + TH cell.

 STEPS INVOLVED IN EXOGEONOUS PATHWAY:

• Peptide generation from internalized molecules (Ag) in endocytic
vesicles.
• Transport of class-II MHC molecule to endocytic vesicles.
• Assembly of peptides with Class-II MHC molecules.
 PEPTIDE GENERATION FROM INTERNALIZED MOLECULES IN
ENDOCYTIC VESICLES:
• Once an antigen is internalized, it is degraded into peptides within
compartments of endocytic processing pathway.
• The endocytic pathway appears to involve three increasingly acidic
compartments, early endosomes (pH 6-6.5), late endosomes or endo-
lysosome (pH 5-6) and lysosomes (pH 4.5-5).
• The internalized antigens move from early to late endosomes and
finally to lysosomes, encountering hydrolytic enzymes and a lower pH
in each compartment.
• It has been suggested that early endosome move from periphery to
inward to become late endosome and finally lysosomes.
 TRANSPORT OF CLASS-II MHC MOLECULE TO ENDOCYTIC
VESICLE:
• When class-II MHC molecules are synthesized within RER, three pairs
of class-II αβ-chains associated with a pre-assembled trimer of a
protein called invariant chain.
• This trimeric protein prevents any endogenously antigen to bind to the
cleft. The invariant chain consists of sorting signals in its cytoplasmic
 ASSEMBLY OF PEPTIDE WITH CLASS-II MHC MOLECULE:

• Class-II MHC-invariant chain complexes are transported from RER through

Golgi complex and Golgi-network and through endocytic compartment,
moving from early endosome to late endosome and finally to lysosome.
• The proteolytic activities increase in each compartment, so the invariant is
slowly degraded.
• However, a short fragment of invariant chain remained termed as CLIP
(Class-II associated invariant chain).
• CLIP physically occupies the peptide binding, cleft of class-II MHC molecule,
presumably preventing any premature binding of antigenic peptides.
• A non-classical class-II MHC molecule known as HLA-DM is required to
catalyze the exchange of CLIP with antigenic peptides.
• The reaction between HLA-DO, which binds to HLA-DM and lessens the
efficiency of the exchange reactions.
• Once a peptide has bound the peptide-class II MHC complex is transported
to the plasma membrane where neutral pH enables the complex to assume
MHC ANTIGENS IN TRANPLANTATION

 In organ transplantation, the MHC genes of the donor and recipient

are important determinants of whether the transplant will be
successful.
 If the MHC genes of the donor and recipient are too different, the
recipient's immune system will recognize the transplanted organ
as foreign and attack it, leading to rejection.
 There are a number of factors that can affect the success of an
organ transplant, including the type of organ being transplanted,
the age and health of the recipient, and the degree of MHC
mismatch between the donor and recipient.
 Rejection responses false into three general categories:
 Chronic rejection
 Acute rejection
 Hyperacute rejection
 HYPERACUTE
REJECTION
 Hyperacute rejection appears within minutes to hours
of placing the transplant when the antigens are
completely unmatched.
 It is caused by the presence of pre-existing antibodies
in the recipient that recognize antigens in the donor
organ.
 It is mediated by humoral system antibodies.
 This type of rejection is seen when a recipient is given
the wrong type of blood. For example, when a person is
given type A blood when he or she is type B.
 ACUTE REJECTION

 This is usually becomes evident within 6 months

of after transplantation.
 Acute graft rejection may be mediated by cellular or
humoral mechanisms.
 The detection of non-self antigens, the formation of
antibodies causes acute rejection.
 By suppressing the immune system, acute rejection
can be treated to some extent and the permanent
damage to the graft can be avoided in some cases.
 CHRONIC REJECTION

 Chronic rejection may develop slowly over a period of

months to a year or so.
 The underlying mechanisms of chronic rejection may be
immunologic or ischemic
 Patients with chronic rejection of renal transplant show
progressive deterioration in renal function as seen by
rising serum creatinine levels.
 There is no cure for chronic rejection except the removal
of graft till date.
 REFERENCE

 Kuby Immunology seventh edition

 [Link]
unology-types-of-graft-and-transplant-rejection/

 [Link]
.com/antigen-processing-and-presentation-cytosolic-
and-endocytic-pathway/