DDS- 2

2017-18

Dr. Shalini Vellasamy

12th March, 2018
At the end of the lesson, the students should be
able to:
1. Explain the genetic organization of the human
MHC, HLA (Human Leukocyte Antigen).
2. Describe the main structural features of class I
and class II MHC gene products.
3. Explain the genetic basis of MHC polymorphism
and its significance for the functioning of the
immune system.
4. Describe the correlation between disease
susceptibility and distinct MHC alleles.
5. List the methods of HLA-typing.
 The major histocompatibility complex (MHC)
was first detected as the genetic locus encoding
the glycoprotein molecules responsible for the
rapid rejection of tissue grafts transplanted
between genetically nonidentical individuals.
(transplantation antigens)
 MHC molecules bind peptide antigens and
present them to T cells. Thus, these
transplantation antigens are responsible for
antigen recognition by the T-cell receptor.
 Histocompatibility refers to the ability to
accept tissue grafts from an unrelated donor.
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 The T-cell receptor is specific for the MHC
molecule.
 If the antigen is presented by another allelic
(variant) form of the MHC molecule in vitro
(normally in an experimental situation), there
is no recognition by the T-cell receptor. This
phenomenon is known as MHC restriction.

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The genetic organization of the MHC:
 In humans, the MHC is also known as Human
Leukocyte antigen (HLA).
 The MHC is a cluster of genes located on
chromosome 6.
 In one individual, all of the principal MHC
gene loci are expressed from both the maternal
and paternal chromosomes.

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 Among the many important genes in the
human MHC, there are three classes of MHC
proteins:
* class I MHC proteins
* class II MHC proteins

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 The number of gene loci (the specific location or position of a

gene, DNA sequence, on a chromosome) for MHC class I and

class II molecules varies between species and
between different haplotypes (a specific group of genes or
alleles that progeny inherited from one parent) within each species.

 The human MHC class I region contains three

principal class I loci – called HLA-A, HLA-B
and HLA-C.
 Each locus encodes the heavy chain of a
classical MHC class I molecule (a
transmembrane glycoprotein) and the whole
region (a non–MHC-encoded polypeptide, β2-
microglobulin). 6/39
 Types of antigens detected by T cells
There are two types of antigen that must be dealt with:
• Endogenous antigens (antigenic peptides from viruses or other pathogens
that inhabit the cell).
• Exogenous antigens (antigenic peptides from
extracellular pathogens that have been taken up by a professional APC).
CD4
APC
CD8
Ag

Exogenous Endogenous Proteins in

microbes/proteins cytosol Ag presented
phagocytosed and Ag to
presented to T cell (MHC I)
T cells (MHC II)
 Class I MHC molecules bind to peptides
derived from cytosolic (intracellular) proteins,
known as endogenous antigens.
 The peptide antigens associated with class I
MHC molecules are recognized by CD8-
positive cytotoxic T lymphocytes.
 Class II MHC molecules bind to peptides
derived from extracellular proteins that have
been brought into the cell by phagocytosis or
endocytosis (exogenous antigens).
 MHC class II–associated peptide antigens are
recognized by CD4- positive helper T cells.
 Class I molecules are found on all nucleated
cells in the body. Exceptions are cells in the
retina and brain.
 The HLA-E, HLA-F, HLA-G and HLA-H
genes also encode MHC class I proteins and
are called class Ib genes.
 They are much less polymorphic than the
HLA-A, -B and -C locus gene products.
 HLA-E and HLA-G gene products can bind to
antigenic peptides, but are involved in
recognition by NK cells.
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The class I MHC locus also includes genes
encoding proteins involved in antigen processing
(eg, transporters associated with antigen
processing [TAPs])
The function of MHC class I molecules is to
present antigens that have entered the cell, such
as viral peptides. Because any cell of the body
may become infected with a virus or intracellular
pathogen, all cells need to sample their internal
molecules and present them at the cell surface to
cytotoxic T cells.
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 Processing of Endogenous Antigens
Cytosolic proteins are Peptide binding to class
MHC class I molecules MHC class I molecules degraded into peptide
I allows it to be
are assembled in the with chaperone fragments by the
released from TAP and
endoplasmic reticulum proteins bind to proteosome.
TAP transports peptides to exported to the cell
(ER) TAP molecules
ER membrane

Cytosolic proteins
can be self proteins
or products of IC
pathogens

• Normal self peptides are constantly being presented on MHC

class I molecules by healthy cells
• Presentation of foreign peptides allows activated T cells to target
infected cells for destruction
 Class II MHC proteins are encoded by the
HLA-D region.
 There are three main families: the -DP, -DQ
and -DR encoded molecules.
 This locus retains control of immune
responsiveness and different allelic forms of
these genes confer striking differences in the
ability to mount an immune response against a
given antigen.

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 The HLA-D locus-encoded proteins are
made up of two noncovalently associated
transmembrane glycoproteins.
 The DR family comprises a single α gene
(DRα) and up to nine β genes (DRβ1–9),
including pseudogenes and several different
gene arrangements occur within the locus.
 The DQ and DP families each have one
expressed gene for α and β chains and an
additional pair of pseudogenes.
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 Unlike class I proteins, they have a restricted
tissue distribution and are chiefly found on
macrophages, dendritic cells, B cells and other
APCs.
 However, their expression on other cells (e.g.
endothelial cells or epithelial cells) is induced
by IFN-γ.
 MHC class II molecules are used by APCs to
present antigens to helper T cells.
Consequently the distribution of class II
molecules is much more limited.
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Processing of Exogenous Antigens
Invariant chain (Ii) binds to Endocytosed proteins are
Invariant
Invariant chain
chain is
is degraded
degraded HLA-DM
HLA-DM molecule
molecule binds
binds to
to
MHC class II molecule in ER degraded in endosomes
in
in endosomes leaving CLIP
endosomes leaving CLIP class
class II,
II, releases
releases CLIP
CLIP so
so
blocking binding of which fuse with vesicles
peptide
peptide blocking
blocking peptide
peptide allowing
allowing exogenous
exogenous
endogenous containing MHC class II
binding
binding groove
groove peptides
peptides to to bind
bind
proteins/peptides molecules

MHC class II
molecules that
bind peptide are
transported to cell
surface

• Proteins from outside the cell are taken up and

degraded in endosomes
• Phagocytosis or receptor mediated endocytosis
• Microbial peptides presented on MHC class II
molecules activate CD4 T cells
 MHC expression
• MHC class I – expressed on all
nucleated cells
• MHC class II – expressed only on
specialised antigen presenting
cells
– Professional APC (DC, Mø)
- Activate naïve T cells
– B cells
- T helper interaction
– Thymic epithelial cells
- T cell development

17
The class III MHC locus encodes:
• complement system molecules
(C4, C2, factor B)
• cytokines (e.g. tumor necrosis factor)
• enzymes
• heat-shock proteins
• other molecules involved in antigen
processing.

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 The genes of the MHC exhibit a remarkable
genetic variability.
 The MHC is polygenic in that there are
several genes for each class of molecule.
 The MHC is also polymorphic. Thus, a large
number of alleles (variants) exist in the
population for each of the genes.
 Each individual inherits a restricted set of
alleles from his or her parent. Sets of MHC
genes tend to be inherited as a block or
haplotype. There are relatively infrequent
cross-over events at this locus. 14/39
 There are a large number of genetic variants
(alleles) at each genetic locus.
 The alleles generally differ from one another
by many (up to 30) amino acid substitutions.
e.g. For some HLA loci, more than 250 alleles
have been identified by serological tests.
 Molecular sequencing has shown a single
serologically defined HLA allele may actually
consist of multiple variants that differ slightly.
 Polymorphism is even greater than that
predicted from serological studies.
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Why the MHC is so polymorphic?
The immune system must handle many different
pathogens. By having several different MHC
molecules, an individual can present a diverse
range of antigens and is therefore likely to be
able to mount an effective immune response.
There is a selective advantage in having different
MHC molecules.
Different pathogens are prevalent in different
areas of the world, so evolutionary pressures
from pathogens will tend to select for different
MHC molecules in different regions. 17/39
MHC class I molecules consist of an MHC
-encoded heavy chain bound to β2-microglobulin.
The class I heavy chain consists of:
• three extracellular domains, designated α1
(N terminal), α2 and α3
• a transmembrane region
• a cytoplasmic tail
β2-Microglobulin is essential for expression of
MHC class I molecules that is also found free in
serum.
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 The products of the MHC class II genes are
HLA-DP, -DQ, and –DR.
 These products are heterodimers of heavy (α)
and light (β) glycoprotein chains and both
chains are encoded in the MHC.
 A number of lines of evidence indicate that
the α and β chains have the same overall
structures.
 An extracellular portion comprising two
domains (α 1 and α 2 or β 1 and β 2) is
connected by a short sequence to a
transmembrane region. 21/39
 The α2 and β2 domains are similar to the
class I α3 domain and β2-microglobulin,
possessing the structural characteristics of
immunoglobulin constant domains.
 Despite the differences in length and
organization of the polypeptide chains, the
overall three-dimensional structure of MHC
class II molecules is very similar to that of
MHC class I molecules.

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 Structure and function of MHC
• MHC I – -chain + 2microglobulin
• MHC II =  chain +  chain
• Structurally similar peptide binding groove
• MHC molecules must bind peptide to be expressed on cell surface

MHC class I MHC class II

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Figure 03-07.   The structure of class I and class II major histocompatibility complex (MHC) molecules.
Antigen recognition by the TCR requires the
antigen to be bound to a specialized antigen
presenting structure known as a major
histocompatibility complex (MHC) molecule.
Unlike immunoglobulins, TCRs recognize
antigen only in the context of a cell–cell
interaction. 25/39

The interaction between peptide

antigen, MHC and the T-cell
receptor is shown.
The Vα and Vβ regions of the
TCR are shown interacting with
the α helices that form the
peptide-binding groove of MHC.
Functions of MHC:
Intracellular antigens are hidden within the cell
and are not available for recognition by antibodies.
How can intracellular antigens be recognized by
extracellular receptors?
 The immune system has solved this problem by
evolving an elegant means of displaying internal
antigens (including those of intracellular
pathogens) on the cell surface, allowing their
recognition by T cells.
 MHC molecules provide a sophisticated
surveillance system for intracellular antigens.
 Proteins within the cell (either produced by the cell or
taken into the cell) are digested into short peptide
fragments.
 Peptides derived from proteins produced within the
cell are displayed on the cell surface through binding to
specialized antigen-presenting molecules termed MHC
class I molecules, which are present on all nucleated
cells of the body.
 Peptides derived from proteins ingested from the
extracellular environment by phagocytosis are
presented by MHC class II molecules, which are
present only on professional antigen presenting cells.
 The peptide–MHC complexes serve as ligands for
TCRs. (lingand is a substance that forms a complex with a biomolecule
to serve a biological purpose) 29/39
An individual’s MHC haplotype affects
susceptibility to disease.
Genetic variations in MHC molecules affect:
 the ability to make immune responses,
including the level of antibody production.
 resistance or susceptibility to infectious
diseases.
 resistance or susceptibility to autoimmune
diseases and allergies.

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• The haplotype HLA-B53 is associated with
protection against childhood malaria, a disease
that is prevalent in equatorial regions.
• The highest frequency of the HLA-B53 allele
is found in Ghana (40%) , China or South
Africa (1-2%).

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HLA types are inherited and some of them are
connected with autoimmune disorders and other
diseases. People with certain HLA antigens are
more likely to develop certain autoimmune
diseases.

(Show-grin's)

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The methods of HLA-typing:
•There are typically three components of HLA
testing used to determine compatibility:
1)HLA typing of donors and recipients
•This step involves identifying HLA alleles.
•HLA matching is important to prevent rejection
•Match the donor and recipient to minimize the
antigenic differences.
•The better the HLA matching of donor and
recipient, the less the strength of rejection.
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• There are many different alleles of the MHC
molecules and one way to reduce the strength
of a rejection response is to match the donor
and recipient so that they share as many alleles
as possible.
• HLA matching is now performed using
molecular techniques, with polymerase chain
reactions (PCR) that are specific for the
different alleles or serological HLA testing.

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2) HLA antibody screening of recipients
 HLA antibody testing is performed on the
recipient to determine if there are any antibodies
present that would target the donated organ or
tissue.

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3) Lymphocyte cross matching (Donor-specific)
 This step occurs after a potential donor has
been identified.
 It helps determine if the intended recipient has
antibodies directed against antigens present on
the donor's lymphocytes.
 Serum from the intended recipient is mixed
with white blood cells (T and B lymphocytes)
from the donor.
Sometimes HLA gene testing is used to aid in
the diagnosis of an autoimmune disease.
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• In humans HLA matching is rarely perfect
between unrelated donors because of the
difficulty in matching all MHC class I and
class II gene loci and the high level of
polymorphism at each locus.
• The importance of HLA matching is not
always crucial.
• HLA matching is very important in bone
marrow transplantation and has a significant
influence on the outcome in kidney
transplantation.
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• For other organs the importance is less clear,
for example:
• In corneal transplantation there is no benefit of
HLA matching;
• For those organs where transplantation is
essential to maintain life (such as heart and
liver) there is no possibility of waiting for a
well-matched organ to become available.

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References:
1. David Male, Jonathan Brostoff, Davis B Roth
and Ivan M Roitt “Immunology” (8th Ed.)
(2013). Elsevier Saunders
2. Geo. F. Brooks, Karen C. Carroll, Janet S.
Butel, Stephen A. Morse and Timothy A.
Mietzner “Jawetz, Melnick & Adelberg’s
Medical Microbiology” (27th Ed.) (2016).
Lange