MAJOR

HISTOCOMPATABILITY
COMPLEX
Dr.T.V.Rao MD

Dr.T.V.Rao MD
1
 MHC - The Major Histocompatibility Complex

• Originally identified in mice as blood cell antigens

by R.A. Gorer and G.D. Snell in 1930s, and defined
on the basis of tissue graft rejection - 1980 Nobel
prize awarded to Snell.
• Work by Rolf Zinkernagel and Peter Doherty in the
1970s revealed that it is the complex of MHC
molecule plus antigen that is recognized by T cells
(MHC restriction of T cell responses) - 1996 Nobel
Prize
• Two classes of MHC molecule: Class I (single MHC
α chain + β2-microglobulin) and Class II (α chain
plus β chain) - more Ig-superfamily
Dr.T.V.Rao MD
members (Ig-C 2
domains)
 Major Histocompatibility complex
(MHC)
• The Major Histocompatibility
complex (MHC) is a large genomic
region or gene family found in most
vertebrates. It is the most gene-dense
region of the mammalian genome and
plays an important role in the immune
system, autoimmunity, and
reproductive success.
Dr.T.V.Rao MD 3
Major Histocompatibility Complex
Definition
• A group of genes that code for cell-
surface Histocompatibility antigens
and are the principal determinants of
tissue type and transplant
compatibility. They are the most
diverse genes in humans and are
used to determine if a sample of DNA
comes from a specific person.
Dr.T.V.Rao MD 4
 MHC
• Genes Of MHC Organized In 3 Classes
– Class I MHC genes
• Glycoproteins expressed on all nucleated cells
• Major function to present processed Ags to TC
– Class II MHC genes
• Glycoproteins expressed on MΦ, B-cells, DCs
• Major function to present processed Ags to TH
– Class III MHC genes
• Products that include secreted proteins that have immune
functions. Ex. Complement system, inflammatory
molecules
 Basis of MHC system
• In man and mouse, as in most species, each
class of MHC is represented by more than one
locus( polygeny), in man these are called HLA
for Human Leucocyte Antigen. The class I loci
are HLA-A,-B and -C and the class II loci HLA-
DR, -DQ and -DP. All the MHC genes map
within a single region of the chromosome (hence
the term Complex); the physical map of the MHC
of man and mouse has been determined by
cloning, mapping and sequencing.

Dr.T.V.Rao MD 6
 Class I, II and III MHC
• Class I MHC Genes Found In Regions A, B and
C In Humans (K and D In Mice)
• Class II MHC Genes Found In Regions DR, DP
and DQ (IA and IE In Mice)
• Class I and Class II MHC Share Structural
Features
– Both involved in APC
• Class III MHC Have No Structural Similarity To
Class I and II
– Ex. TNF, heat shock proteins, complement
components
 Composition of Class I
Molecules
• Class I molecules are composed of two
polypeptide chains; one encoded by the BCA
region and another (ß2-microglobulin) that is
encoded elsewhere. The MHC-encoded
polypeptide is about 350 amino acids long and
glycosylated, giving a total molecular weight of
about 45 kDa. This polypeptide folds into three
separate domains called alpha-1, alpha-2 and
alpha-3. ß2-microglobulin is a 12 kDa
polypeptide that is non-covalently associated
with the alpha-3 domain.

Dr.T.V.Rao MD 8
 Class II molecules
• MHC (major Histocompatability
complex) Class II molecules are
found only on a few specialized
cell types, including
macrophages, dendritic cells and
B cells, all of which are
professional antigen-presenting
cells (APCs).
Dr.T.V.Rao MD 9
 Humans are very
Diversified MHC Plays a Great Role
The loci which specify MHC’s are
polymorphic.
Many alleles may exist at a locus:

HLA A locus has ~60 alleles

HLA B locus ~110 alleles
HLA C locus ~40 alleles

The high level of allelism creates diversity

within a species (thus restricting allografting)
but does not produce diversity within an
individual
Dr.T.V.Rao MD 10
 What is polygenecity?
• Humans have DP, DQ, and DR “regions”
specifying α and β chains of MHC-II.
• Why are these called “regions”?
 MHC Molecules Control the
Immune response
• The Major Histocompatibility Complex
(MHC) is a set of molecules displayed on
cell surfaces that are responsible for
lymphocyte recognition and "antigen
presentation". The MHC molecules control
the immune response through recognition
of "self" and "non-self" and, consequently,
serve as targets in transplantation
rejection
Dr.T.V.Rao MD 12
 Peptide Binding to MHC Molecules: Class I vs. II

Class I Class II
• Generated by proteasome (cytosol) • Generated in endosomes/lysosomes

• Transported to ER by TAP1/2 • Bind in specialized vessicles (MIIC)

• Restricted length, 8-9 residues • Any length, extend from MHC

• Binding energy from N, C termini • Binding energy from backbone

• “Anchor” residues fill specific pockets • “Anchor” residues fill specific pockets

Dr.T.V.Rao MD 13
 MHC- I MHC-II
MHC-I

Heavy chain
(alpha) and
“microglobulin”
(beta two)

Heavy is 45
kilodaltons, has
three domains +
a
transmembrane
component (40
aa) + a cyto-
plasmic tail (30
aa)

The three alpha

domains are
called: α1, α2, &
α3
α1 and α2 interact to MHC-II
present
processed Ag An alpha and beta chain, 33 kDA and
Process Ag is 28 kDa, respecitvely.
optimally a
nonomer Chains are non-covalently associated.
Each chain has two domains.
Microglobulin (12 kDa) associates non-covalently with α3 α1-β1 interact to present processed Ag
Microglobulin and α3 are part of immunoglobulin Processed Ag is optimally 13-18 aa
superfamily

Microglobulin is the only member of the superfamily that α2 & β2 are part of immunoglobulin
does not have a component linking it to a membrane super family
 The Major Histocompatibility Complex
• MHC Class I and Class II genes are encoded in a large
(>4 Mb, >200 genes) tightly linked gene cluster: the
MHC
• The MHC is similarly organized in mice (Ch17) and
humans (Ch6) - syntenic
• Known in humans as HLA (human leukocyte antigen)
genes; in mouse as H-2 (histocompatibility group 2)
genes
• Genes are highly polymorphic (many alleles)
• Class I: HLA-A, -B, -C (human); H-2K, -2D, -2L
(mouse)
• Class II: HLA-DP, -DQ, -DR (human), I-A, I-E (mouse)
Dr.T.V.Rao MD 15
 Human
MHC class I:
HLA-A, -B, -C
MHC class II:
HLA-DR, -DP, -DQ

Mouse
MHC class I:
H2-K, -D, -L
MHC class II
H2-A, -E (I-A, I-E)

β 2-microglobulin is located on a different

chromosome.
Dr.T.V.Rao MD 16
 MHC is represented …
• In man and mouse, as in most species, each
class of MHC is represented by more than one
locus(polygeny), in man these are called HLA for
Human Leucocyte Antigen. The class I loci are
HLA-A,-B and -C and the class II loci HLA-DR, -
DQ and -DP. All the MHC genes map within a
single region of the chromosome (hence the
term Complex); the physical map of the MHC
of man and mouse has been determined by
cloning, mapping and sequencing.

Dr.T.V.Rao MD 18
Class I and II MHC

Dr.T.V.Rao MD 19
MHC- I & MHC-II

Dr.T.V.Rao MD 20
Dr.T.V.Rao MD 21
 Class I And II Specificity
• Several Hundred Allelic Variants Have Been
Identified In Humans
• However, up to 6 MHC I And 12 MHC II Molecules
Are Expressed In An Individual
• Enormous Number Of Peptides Needs To Be
Presented Using These MHC Molecules
• To Achieve This Task MHC Molecules Are Not Very
Specific For Peptides (Unlike TCR and BCR)
• Promiscuous Binding Occurs
– A peptide can bind a number of MHC
– An MHC molecule can bind numerous peptides
 Class I And II Diversity And
Polymorphism
• MHC Is One Of The Most Polymorphic
Complexes Known
• Alleles Can Differ Up To 20 a/a
• Class I Alleles In Humans: 240 A, 470 B, 110
C
• Class II Alleles In Humans: HLA-DR 350 β, 2
α!
• HLA-DR
β genes vary from 2-9 in different individuals!!!,
– 1 α gene (α can combine with all β products
increasing number of APC molecules)
 MHC I and II belong to
Immunoglobulin Supergene Family
• The Class I and Class
II MHC molecules
belong to a group of
molecules known as
the Immunoglobulin
Supergene Family,
which includes
immunoglobulin's, T-
cell receptors, CD4,
CD8, and others

Dr.T.V.Rao MD 24
MHC location on Chromosome 6
• The major
Histocompatibility
complex is encoded by
several genes located on
human chromosome 6.
Class I molecules are
encoded by the BCA
region while class II
molecules are encoded
by the D region. A region
between these two on
chromosome 6 encodes
class III molecules,
including some
complement components.
Dr.T.V.Rao MD 25
 MHC Expression
• Expression Is Regulated By Many Cytokines
– IFNα, IFNβ, IFNγ and TNF Increase MHC
expression
• Transcription Factors That Increase MHC gene
Expression
– CIITA (Trans activator), RFX (Trans activator)
• Some Viruses Decrease MHC Expression
– CMV, HBV, Ad12
• Reduction Of MHC May Allow For Immune
System Evasion
 MHC system important in
Rejection of Transplants
• Gene products encoded in the Major
Histocompatibility Complex (MHC)
were first identified as being important
in rejection of transplanted tissues.
Furthermore, genes in the MHC were
found to be highly polymorphic (i.e. in
the population there were many
different allelic forms of the genes)
Dr.T.V.Rao MD 27
 Class I MHC proteins
• Class I MHC proteins (found on all nucleated cell
surfaces) present antigens to cytotoxic T lymphocytes
(CTLs) . Most CTLs possess both T-cell receptors (TCR)
and CD8 molecules On their surfaces. These TCRs are
able to recognize peptides when they are expressed in
complexes with MHC Class I molecules. For the TCR to
bind a peptide-MHC complex two conditions must be
met. Firstly, the TCR must have a structure which allows
it to bind the peptide-MHC complex. Secondly, the
accessory molecule CD8, must bind to the alpha-3
domain of the MHC Class I molecule. Due to genetic
recombination events each CTL expresses a unique
TCR which only binds a specific MHC-peptide complex

Dr.T.V.Rao MD 28
 Class I MHC molecules
• Class I MHC molecules
are composed of two
polypeptide chains, a
long α chain and a short
β chain called β2-
microglobulin (figure 2).
The α chain has four
regions. First, a
cytoplasmic region,
containing sites for
phosphorylation and
binding to cytoskeletal
elements

Dr.T.V.Rao MD 29
Class II MHC molecules
• Class II MHC molecules
are composed of two
polypeptide chains an α
and a β chain of
approximately equal
length (Figure 6). Both
chains have four regions:
first, a cytoplasmic region
containing sites for
phosphorylation and
binding to cytoskeletal
elements;

Dr.T.V.Rao MD 30
MHC II are Complex in
Organization
• Within the MHC there are
5 loci that encode class II
molecules, each of which
contains a gene for an α
chain and at least one
gene for a β chain. The
loci are designated as
HLA-DP, HLA –DQ, HLA-
DR, HLA-DM, and HLA-
DO. Among these, HLA-
DP, HLA –DQ, and HLA-
DR are the most
important and are most
polymorphic..

Dr.T.V.Rao MD 31
 MHC is highly polymorphic
• The MHC is highly polymorphic. There are a
large number of genetic variants (alleles) at
each genetic locus. Crucially many of these
alleles are represented at significant frequency
(> 1%) in the population, and in addition the
alleles generally differ from one another by many
(up to 30) amino acid substitutions. For example
in humans there are more than 200 alleles
described at some MHC loci. Such a remarkable
degree of polymorphism implies a selective
pressure to establish and maintain it.

Dr.T.V.Rao MD 32
MHC Polymorphism

Dr.T.V.Rao MD 33
 MHC Polymorphism
• MHC Class I and Class II molecules have
many alleles
• Diversity ensures that a wide range of
peptides can be presented within the
population (even if a much more limited
set is presented by any individual)
• For Class II, both α and β chains are
polymorphic (except DRα in humans and
Eα in mice), adding more diversity
Dr.T.V.Rao MD 34
Inheritance Of HLA Haplotypes
 MHC Polymorphism
4)Collection of Class I +
Class II alleles on one
chromosome is called
the haplotype
5)MHC molecules are
expressed co-
dominantly: alleles
from both chromosomes
are expressed in each
cell

Dr.T.V.Rao MD 36
 Biological role of the MHC

• The products of the MHC

play a fundamental role in
regulating immune
responses. T cells must
recognise antigen as a
complex with MHC
molecules. This requires
antigen to be processed
by unfolding and
proteolytic digestion
before it complexes with
the MHC molecule

Dr.T.V.Rao MD 37
Biological role of the MHC

• Once formed the complex

of antigenic peptide and
MHC are generally very
stable (half life ~ 24hrs).
Thus the biological role of
MHC proteins is to bind
small peptides and to
"present" these at the cell
surface for the inspection
of T cell antigen
receptors.

Dr.T.V.Rao MD 38
 MHC-Linked Diseases
• Defects in MHC gene expression
lead to immunodeficiencies (MHC
molecules are required for both T
cell development and activation)
• Some MHC alleles are associated
with susceptibility or resistance to
autoimmune diseases
Dr.T.V.Rao MD 39
 MHC-Linked Immunodeficiencies
Bare Lymphocyte Syndromes lead to
loss of MHC molecule expression:
• Defects in TAP genes prevent MHC Class I
protein surface expression (even though
MHC proteins are normal), so no CD8+ T
cells - surprisingly mild immunodeficiency
(respiratory and skin infections)
• Defects in TF’s controlling Class II gene
expression (CIITA, RFXANK, RFX5,
RFXAP) block CD4+ T cell development -
result in SCID (severe combined
immunodeficiency)Dr.T.V.Rao MD 40
• Programme Created by
Dr.T.V.Rao MD for Medical
Students in the Developing
World
• Email
• doctortvrao@gmail.com
Dr.T.V.Rao MD 42