Major

ajo Histocompatibility
stoco pat b ty Complex
Co p e
encoded proteins
Dipankar Nandi

Department of Biochemistry
Indian Institute of Science (IISc)
Bangalore

Email: nandi@iisc.ac.in
Tel: 080-2293-3051

g , 2019
28 & 29 August,
Important concepts about MHC and relationship
with transplantation
 1940’s : Graft rejection
Transplantation antigen (histocompatibility antigen)
Antigens which cause immune response to the graft and
determine the survival of the graft.
Alloantigens specific for each individual .
History off transplantation is often
f linked
k d to MHC!

 MHC ((Major j histocompatibility

p y complex)
p )
A large cluster of linked genes located on Chromosome 6 in
humans or Chromosome 17 in mice that encodes proteins
responsible for immune responsiveness,
responsiveness e.g.
e g transplant rejection,
rejection
immune response, susceptibility to autoimmune diseases, etc.

How??
H
Encode proteins that present peptides to T cells.
 Peter Medawar
The Nobel Prize in Physiology or Medicine- 1960

World War II at Burns Unit of the

Glasgow Royal Infirmary in Scotland

Laws of transplantation

Acquired
A i d tolerance
l generation
i and
d
mechanisms

Medawar,, P.B. ((1951).

) The Technique
q of Free Skin
Grafting in Mammals. J Exp Biol. 28: 385-402.
 Brief history of MHC research using mice
Abbie Lathrop: retired school teacher who bred and supplied mice
for research. Several mice that are commonly used today originated
from her farm,
farm e.g.
e g C57BL/6 = female number 57.
57

William Castle of the Bussey Institute at Harvard University initiated

research
h on mice
i as a model d l system. Mice
Mi were supplied li d by
b A. A
Lathrop’s farm. Trained several scientists, e.g. C. Little, G. Snell, etc,
who made major contributions to mouse genetics, immunology, etc.

Clarence Little developed “inbred” strains of mice, e.g. C57BL/6,

BALB/c (Bagg
(Bagg’ss albino),
albino) etc.
Founded the Jackson Lab, Bar Harbor, Maine – which has pioneered
research in mice, large collection of mice mutants (e.g. Fas, FasL, Leptin
k k t [ob
knockout [ b-/-
/ ] and
d Leptin
L pti receptor k k t[db-/-
pt knockout[db / ],
] etc)
t ) andd lead
l d the
th mouse
genome project.
 Brief history of MHC research using mice
G. Snell: Ability to reject tumors in mice was genetically determined.
In collaboration with P. Gorer, discovered H-2 (mouse MHC). H-2
was the second genetic locus involved in control of expression of
antigens important for transplantation.
Developed the system of developing congenic mice (different
chromosomal segment)

B. Benacerraf: Role of IR genes, ability to respond to an antigen was

determined by IR genes (MHC class II).

1980 Nobel prize awarded to Snell, Benacerraf and Dausset.

Benaceraff, Snell and Dausset
The Nobel Prize in Physiology or Medicine - 1980

Baruj Benacerraf George D. Snell Jean Dausset

Ir g
genes Congenic strains yp g
HLA typing
Mapping H2 genes Organ transplants
MHC classII

Determine Ab
responses
 The Jackson Laboratory
 Abbie Lathrop

 Clarence Little

 Centre of Mouse Genetics

 Two Fires

 Fas (lpr), FasL (gld),

Leptin (ob), Leptin
p
receptor ((db).
)

 Mouse Genome Project

The Jackson Laboratory
Bar Harbor, Maine
A BLACK SPOT!

1972: the case of W. T. Summerlin,

skin transplant with black ink.
MHC - The
Th Major
M j r Histocompatibility
Hi t p tibilit Complex
C pl

Two classes of MHC molecule:

 Class I ((single
g MHC α chain + β2 microglobulin)
g )

 Class II (α chain plus β chain) – two chains

 Structure of MHC
MHC class I Molecule MHC class II Molecule
Peptide
P ptid binding
bi di Peptide binding
groove groove
α2 α1 β1 α1

α3 β2m α2
β2
Cellular T cell Immunity and Organ Transplants
T cells are activated with antigens (MHC + peptide)

MHC-
peptide
T cell
APC
TCR

 IL2 Cytokine production.

 T cell proliferation.

 Killing by CTLs.
MHC the Major Histocompatibility Complex
MHC,

MHC class I MHC class II

Human HLA A HLA

HLA-A, HLA-B,
B HLA-C
HLA C HLA-DR,
HLA DR HLA-DP,
HLA DP HLA-DQ
HLA DQ

Mouse H2-K, H2-D, H2-L H2-IA, H2-IE

β2-microglobulin
l b l is llocated
d on a ddifferent
ff chromosome.
h
MHC, the Major Histocompatibility Complex

MHC
C Cl
Class I MHC
C Cl
Class II
HLA-A, HLA-B, HLA-DP, HLA-DQ,
Nomenclature
HLA C
HLA-C HLA-DR
A

Macrophages,
p g , B-cells,,
All nucleated Dendritic cells,
Found on
somatic cells Langerhans cells in skin,
activated human T cells

Recognized by CD8+ TC cells CD4+ TH cells

MHC, the Major Histocompatibility Complex

MHC Class I MHC Class II

Presentation of Ag to Presentation of Ag to
TC cells leading to TH cells which secrete
elimination of tumor or cytokines
y and provide
p
infected host cell. help.
 HLA
Chromosome 6 MOG

HLA-A
6p (short arm)
Class I

HLA-C
HLA-B

TNF
Hsp
Class III
Cp
6q (long arm)

HLA-DR
HLA-DQ

Class II
HLA-DP

TAPasin
 THE MHC GENE COMPLEX

The MHC complex contains a number of genes that control

several antigens, can be divided into three major classes:

 Class I Expressed on cell surface

 Class II
 Class III. Present in body fluids
(e g C4 C2
(e.g.C4, C2, factor B,
B TNF
MHC class I & MHC class II
Encodes for peptide-binding proteins known as MHC class I & II respectively.
MHC I, that are expressed on most nucleated cells.

MHC class I is composed of a heavy chain that is associated with β2-

microglobulin and a peptide. MHC class I molecules present peptides to CD8+
T-cytotoxic cells.

MHC class II molecules are recognized by CD4+ T helper cells and are
composed of a peptide and two proteins, α and β chains. MHC class II
molecules
l l are expressedd only
l on APCs,
APC e.g. dendritic
d d i i cells,
ll macrophages,
h etc

MHC class III

Encodes different immune components, e.g. complement components
(e.g. C2, C4, factor B, cytokines (e.g., TNF ß) and also heat shock proteins.
 Human MHC Locus

The human MHC is located on chromosome 6.

Class I MHC:

 Contains three major loci, B, C and A and undefined minor loci.

 Each major locus codes for a polypeptide.
polypeptide
 The α-chain is polymorphic (has many alleles).
 β-2 microglobulin (beta-chain) is encoded by a gene outside the MHC complex.
 Without the ββ-2 microglobulin,
g , the class I antigen
g will not be expressed
p leadingg
to deficiency of cytotoxic CD8+ T cells.

Class II MHC

 The class II gene complex also contains three loci, DP, DQ and DR.
 Each of these loci codes for one α and one β chain polypeptide.
p yp p
 Key features of MHC
Binds & presents peptides to T cells; peptide stabilizes cell
surface MHC interaction.
interaction

 Genetic localization.

. Polygenic (several genes) & polymorphic (different alleles)

Co-dominant expression (see Fig 8-12 in Kuby).

 IFN/ cytokine inducible

 World : Proteins

Continents : Superfamily

Countries : Ig TFs

Districts : Ig TCR MHC Adhesion

Village : Classical Non-classical

Class I Class II

Haplotype used HLA-A, HLA-B, HLA-C

in mice
Polymorphisms
H2k, H2d, H2b HLA-B27-01, HLA-B27-05
 HUMAN MHC (HLA)
HLA-A + 2M

HLA-B + 2M
HLA-C + 2M
Total SIX different MHC class I proteins (HC + 2M) in a
HUMAN cell – all nucleated cells express
p MHC class I molecules
that are “co-dominantly” expressed

HLA-DPalpha & DPbeta

HLA-DQalpha
HLA DQalpha & DQbeta
HLA-DRalpha & DRbeta
Totall SIX different
T diff MHC class
l II proteins
i ((alpha
l h &b beta chain)
h i ) are
“co-dominantly” expressed in an antigen presenting cell – note
MHC class II expression is in APCs and not on all cells.
 MOUSE MHC (H-2)
H2-K + 2M H2-IEalpha & IEbeta
H2-D + 
2M H2-IAalpha
p & IAbeta
H2-L + 2M

Theoretically, SIX different MHC class I and FOUR types of

p
MHC class II molecules can be expressed – onlyy in outbred
mice or recombinants.

In a haplotype, e.g. H2k or H2b, only three different MHC class

I and two MHC class II molecules will be expressed.
Why? Genomes of males & females same in inbred mice!
Differences: Antigen Processing and Antigen Presentation
MHC molecules bind peptides inside the cell and present them to
T cells on the cell surface

We need to consider at least two scenarios

I) II)

MHC (self) MHC (self)

+ +

Self peptides peptides derived from microbes or

tumor antigens or non-self peptides

No immune response Immune response

by T cells (majority) by T cells (upon infection, cancer etc)

TOLERANCE IMMUNE RESPONSE

How does one experimentally demonstrate this??

 Zinkernagel and Doherty experiment

Note H2K and H2b are

alleles, i.e. different
forms of MHC!

H2k (self H2b (self MHC)

MHC) +
+ LCM virus
LCM virus
Wrong combo
Right combo
Why?
Why?
 Doherty and Zinkernagel
The Nobel Prize in Physiology or Medicine -1996
1996
Peter C. Doherty
Recognition of virus-infected cells by the
immune system.
system

Proposed and experimentally demonstrated

the “altered
altered self ” hypothesis:

Self = MHC

Alteration of MHC occurs upon infection.

Altered Self = MHC + foreign peptide

derived from an invading pathogen or a
tumour.
tumour

Rolf M. Zinkernagel
MHC restriction

Note right MHC and right antigenic peptide is required

for recognition by appropriate T cell receptor (TCR)

Self MHC (selected) + peptide derived from pathogen

This was the conclusion of the Zinkenagel and

Doherty’ss experiments on “altered
Doherty altered self hypothesis
hypothesis” –
self is MHC which gets altered upon binding of
pathogen/tumor encoded peptide
Mice (H2d) Mice (H2d)

Inject with APCs Inject with heat killed E. coli

(H2b)
After few weeks
weeks, isolate T
After few weeks, cells from mice and study
isolate T cells from responses with
mice and study
responses with APC (H2b) without infection

APC (H2b) APC (H2b) with infection

or APC (H2d) without infection

APC (H2d) APC (H2d) with

ith iinfection
f ti

ALLO- Ag-specific responses

RESPONSES
MHC CLASS I Dependent Antigen Presentation
Peptide generation in CYTOSOL

Peptide translocation into ER

Peptide binding to MHC class I

Trafficking to the cell surface

Intra cellular antigen

ER (
(e.g. viruses)
ir )
Proteolysis
Nucleus

MHC I Peptide binding to MHC in ER

Golgi
MHC I on cell surface
 MHC CLASS II Dependent Antigen Presentation

Extra cellular antigen

is endoc
endocytosed
tosed and degraded
in lysosome-like vesicles

Nucleus ER
Nucleus
Peptide binding
in MHC II

MHC II presents MHC II in

Golgi
peptide at cell surface vesicle
MHC is closely associated with
1) T cell activation in the periphery,
periphery ee.g.
g lymph nodes
nodes, spleen
spleen, etc
etc.
2) T cell education in thymus.
This is an important concept!

Defective MHC I expression

p Defective MHC II expression
Low expression of MHC class I.
Low numbers of CD8+ T cells.
cells
Recurrent severe pulmonary
infections. Disease severity
v y withw
respect to survival varies in
patients.
p
No HLA-DP, -DQ and -DR
expression.
Low numbers of CD4+ but
normal to elevated numbers of
CD8+ T cells.
ll
Often associated with viral
infections (Coxsackie,
(Coxsackie herpes),
herpes) oral
candidiasis, bacterial pneumocystis
and septicemia.
 MHC Li k d Di
MHC-Linked Diseases

 Defects
D f in
i MHC gene expression
i lead
l d to immunodeficiencies
i d fi i i
(MHC molecules are required for both T cell development and
activation).

 Some MHC alleles are associated with susceptibility or

resistance to autoimmune diseases.
 Associations of HLA type and
susceptibility to autoimmune disease.
disease

Associations of HLA serotype

yp with susceptibility
p y to
autoimmune disease
Disease HLA Allele Relative risk
Ankylosing spondylities B27 87.4
Acute anterior uveitis B27 10
Good pasture’s syndrome DR2 15.9
Multiple sclerosis DR2 4.8
G
Graves’ ’ disease
di DR3 37
3.7
Myasthenia gravis DR3 2.5
 Associations of HLA serotype
yp and
susceptibility to autoimmune disease
Associations of HLA serotype with susceptibility to
autoimmune disease
Disease HLA Allele Relative risk
Systemic lupus DR3 5.8
erythematosus
Type –I insulin DR3/DR4 ~ 25
dependent Diabetes
Rheumatoid arthritis DR4 4.2
MHC U
U-tube:
tube: https://youtu.be/yDAGxVxY
https://youtu be/yDAGxVxY-L8
L8

Al
Also, d
do read
d up on th
the GENOGRAPHIC PROJECT

https://genographic.nationalgeographic.com/about/
p g g p g g p

https://en.wikipedia.org/wiki/Genographic_Project

The Genographic Project, launched on 13 April

2005 by the National Geographic Society and IBM,
is a multi-year
lti genetic
ti anthropology
th l study
st d th
thatt aims
i s
to map historical human migration patterns by
g and analyzing
collecting y g DNA samples
p from
hundreds of thousands of people from around the
world
KEY CONCEPTS:
MHC: HLA ((human),
) H-2 mouse))
chromosomal localization
different classes (class I, II, III) and functions
key features
MHC class I and MHC class II
Zinkernagel & Doherty’s expt to show MHC restriction
51Cr-release assay for cytotoxicity
Antigen processing experiment
Differences in the antigen processing pathways for MHC class I and
MHC class II – why?y How does it benefit the immune system?
y

TERMS: Codminance, inbred strains of mice, polygenic, polymorphisms, antigen

processing, antigen presentation, HLA, H-2, haplotype, intracellular antigens,
extracellular antigens