Beruflich Dokumente
Kultur Dokumente
Presentation
Chapter 8
MAJOR HISTOCOMPATIBILITY
COMPLEX ( MHC )
1) Role in organ transplantation
2) Role in immune response
Vaccine immunity
Infectious diseases
MHC MOLECULES
Peptide
Binding
Cleft or
Groove
MHC class I
Peptide
Foreign Peptides
Virus
Bacteria
Parasite
Fungi
Allergen
Transplant
MHC class II
Peptide
Binding
Cleft or
Groove
Self Peptides
Cryptic self peptides
(autoimmune Ag)
Altered self (cancer)
Cell Membrane
MHC Properties
Are the most polymorphic genes
(alternative forms of a gene/ allele) in
genome (population)
Also polygenic (several genes with similar,
but not identical structures and functions),
on individual level
The set of MHC alleles on each
chromosome is a haplotype (HLA-A2,HLAB5, HLA-DR7,etc
Each person gets 2 haplotypes- one from
mom and the other from dad
Chromosome 17
Glycoprotein (Gp)
or Protein (P)
(Human leukocyte antigen complex)
Chr. 6
Gp
or P
MHC Properties
Much of our initial knowledge of MHC came
from mice (KO mice-MHCIKO mice)
20 inter-matings make identical (syngenic)
mice
Syngeneic inbred mice have a single haplotype
Allogeneic, (distinct strains of mice- eg. Balb/c
versus C57BL/6) may have differing
haplotypes)
Congeneic strains identical except at 1 locus
Are people syngeneic or allogeneic?
11
MHC Properties
MHC molecules have been found in every
mammalian species examined
MHC alleles are Co-dominantly expressed
In humans, each HLA allele is given a number
designation
In mice, each H-2 allele is given a letter
designation
12
MHC Properties
MHC molecules consist of extracellular peptide
binding cleft (groove), Ig-like domain, a
transmembrane domain, and cytoplasmic region
Polymorphic AA of MHC located in and adjacent
to peptide binding groove
Consist of paired a helices and a floor of 8
stranded b pleated sheets
Non-polymorphic Ig domains of MHC interact
with CD4 and CD8 molecules
13
MHC
Peptides that bind to a given MHC share
structural features that promote interaction
Association of antigenic peptide / MHC has
slow off rate
MHC molecules dont discriminate between
foreign and self peptides
Hydrophobic interactions, hydrogen bonding
and charge interactions play large role in
association between peptide and MHC
14
MHC Class I
Consist of an a chain and a Non-MHC encoded
subunit (b2-Microglobulin)
Present endogenously derived peptide to Tc
Binds peptides 8-11 amino acids in length
This means that native proteins have to be
processed prior to binding
15
MHC Class II
Consist of MHC encoded a chain and b chain
subunits
Present exogenously derived peptide to Th
Binds peptides 13-20 amino acids in length
Again, the native proteins have to be
processed prior to binding
16
Class III
Within Class II
DP2, DP2 - pseudogenes
DQ2, DQ2, DQ3 Not expressed
18
Question:
Female Mouse of MHC Haplotype H-2b
(C57BR) is mated with male Mouse of MHC
Haplotype H-2d (Balb/c).
What would be the MHC haplotype of the
progeny?
Would transplanted tissue from the mom be
accepted or rejected by the progeny?
Would transplanted tissue from the progeny
be accepted or rejected by the mom?
Would transplanted tissue from the dad be
accepted by the mom?
19
20
21
22
MHC of an individual
2 MHC Haplotypes: one from mom and dad
Each haplotype has one HLA-A, B, C
Each haplotype has at least one HLA-DR, DP,
DQ, however there are multiple alpha and
beta chains
Therefore it is estimated that each individual
has 6 MHC I and up to 12 MHC II
23
MHC
Analysis of human MHC alleles in 2006 have
revealed 370 A alleles, 660 B alleles, 190 C
alleles.
Numbers likely to be significantly larger as
data determined from European descent. In
fact, many non-European groups cannot be
typed using available MHC serological typing
reagents
24
DQ
DR
b a
b a
b1 a
A
Polygeny
DP
b a
DQ
b a
DR
b1 a
DP
b a
DQ
b a
DR
b1 a
A
Variant alleles
polymorphism
HAPLOTYPE 1
HAPLOTYPE 2
Additional set
of variant alleles
on second
chromosome
SOME 6
mere
HLA HAPLOTYPE
DP DQ DR
Class III
B C
Class I
DR55
DQ63
DR
45
DQ
33
B7 C9
DQ36
DR54
DP
11
DQ66
DR44
81
(2 x A) + (2 x B) + (2 x C) = 6 MHC-I
8
APC
(4 x DR) + (4 x DP) + (4 x DQ) = 12 MHC-II
A11
INHERITANCE OF
HLA
HAPLOTYPES
DP-1,2
DQ-3,4
DR-5,6
B-7,8
C-9,10
A-11,12
DP-9,8
DQ-7,6
DR-5,4
B-3,2
C-1,8
A-9,10
Parents
DP
DP
DQ
DQ
DR
DR
B C
B C
Children
DP
DQ
DR
B C
DP
DQ
DR
B C
What is a haplotype ?
A set of linked MHC alleles present on
one parental chromosome.
Thus, an individual has two haplotypes.
DP-1,9
DQ-3,7
DR-5,5
B-7,3
C-9,1
A-11,9
DP-1,8
DQ-3,6
DR-5,4
B-7,2
C-9,8
A-11,10
DP-2,8
DQ-4,6
DR-6,4
B-8,2
C-10,8
A-12,10
DP-2,9
DQ-4,7
DR-6,5
B-8,3
C-10,10
A-12,9
DP
DQ
DR
B C
DP
DQ
DR
B C
DP
DQ
DR
B C
DP
DQ
DR
B C
DP
DQ
DR
B C
DP
DQ
DR
B C
DP
DQ
DR
B C
DP
DQ
DR
B C
Papain
COOH
Description
Peptide-binding domain
Peptide-binding cleft
General bound peptide size
Peptide binding motifs
COOH
COOH
MHC-I Molecules
MHC-II Molecules
1 / 2
1 / 1
MHC-I MOLECULE:
SCHEMATIC DIAGRAM & DESCRIPTION
MHC-encoded a-chain of 45kDa
a-chain anchored to the cell membrane
a2
a1
a3
b2-m
MHC-II MOLECULE:
SCHEMATIC DIAGRAM & DESCRIPTION
MHC-encoded, a-chain of 33kDa
and a b-chain of 28kDa
a and b chains anchored to the cell membrane
b1
b2
a1
a2
No b2-microglobulin
Peptide antigen in a groove formed from a pair of
a1/1-helixes on a floor of anti-parallel b strands
a2 & b2 domains have structural & amino acid
sequence
homology with Ig constant domains - members of
Ig GENE SUPERFAMILY
Peptide
a-chain
b-chain
a-chain
a-chain
Peptide
b2-m
Peptide
b-chain
33
MHC-BINDING PEPTIDES
Each human usually expresses:
6 types of MHC class I (A, B, C) and
at least 12 types of MHC class II (DR, DP, DQ)
Total of 18 MHC molecules per person
1,000,000,000,000,000 (1x1015).
Each of which may potentially recognizes a different peptide antigen.
Floppy
Compact
P S S
P9
I
A K
I
MHC-I molecule
P3
S A
P9
K S I P S
Y
I
MHC-I molecule
HUMAN MHC:
Number of HLA Class-I Alleles:
660 HLA-B > 370 HLA-A > 190 HLA-C
4.6x107 MHC-I combinations
HLA Class-II for Antigen Presentation:
3 HLA-DRA, 480 HLA-DRB,
(15) HLA-DPA1, 118 HLA-DPB1,
28 HLA-DQA1, 62 HLA-DQB1
8x1011 MHC-II combinations
4x1019 MHC-I & -II combinations
Signal Peptide
EXTRACELLULAR REGION
A
HLA-A*0201
W
L
I
L
L
L
S
F
hK
L
G
E
L
V
P
V
P
V
P
V
H
F
hY
G
V
V
Y
HLA-A3
R
I
R
L
L
L
R
R
R
P
G
A
G
S
E
G
V
A
K
A
G
K
H
- V
K
K
K
HLA-A*6801
K
E
A
T
V
V
G
hA
A
G
P
A
P
P
V
I
E
A
Y
Y
A
K
H
- R
R
R
R
HLA-B7
G
I
P
P
P
P
G
Q
hP
P
C
I
Q
R
F
P
L
I
G
T
R
P
P
hR
L
L
L
HLA-B27
R
R
G
R
R - R
V
I
I
K
D
K
E
K
E
V
P
I
V
I
V
K
L
- K
K
K
K
Cytoplasmic Domain
BINDING GROOVE
Pocket
Pocket
Pocket
Pocket
Pocket
Pocket
A
B
C
D
E
F
PEPTIDES
P1 P2 P3 P4 P5 P6 P7 P8 P9
Transmembrane Domain
NH3+
P4 P5
P8 P
P1
P2 P3
Pockets
of an HLA A B
molecule Adapted from
h = hydrophobic aa
P
6
D
P7
P9
MHC
XX
Pathogen that
evades MHC X
Survival of
individual
threatened
Pathogen that
evades
MHC X
but has
sequences
that bind to
MHC Y
MHC
XX
MHC
YY
MHC
XY
Impact on the
individual depends
upon genotype
Population survives
MHC
YY
MHC
XY
Survival of individual
threatened
each individual had 6 types (HLA-A, -B, -C, -DR, -DP, -DQ) of
MHC the alleles of each MHC type were randomly distributed in the population
any of the 1,900 alleles could be present with any other allele
CAU
AFR
ASI
HLA-A1
15.18
5.72
4.48
HLA- A2
28.65
18.88
24.63
HLA- A3
13.38
8.44
2.64
HLA- A28
4.46
9.92
1.76
HLA- A36
0.02
1.88
0.01
MHC-RESTRICTION IN
ANTIGEN PROCESSING AND PRESENTATION
Fixation blocks
metabolic
activity.
No Ag
Processing or
Presentation
Ag Presentation
Ag Processing
Ag Peptide
Binding
Uninfected
Ag Presentation
MHC restriction:
Effector (Tc) cells and target cell
need to be MHC identical.
MHC class I
T cells
+++
- / + (activation)
B cells
Macrophages
Dendritic cells
+++
+++
+++
++
++ (activation)
+++
Neutrophils
Hepatocytes
Kidney
Brain
Erythrocytes
+
+
+
+++
+
+
+
-
MHC class II
+++ (activation)
++ (activation)
human + ++
PROTEIN
FUNCTION
HLA - A
HLA - B
MHC-I
(all nucleated cells)
HLA - C
HLA - DR
HLA - DQ
MHC-II
(APC)
HLA - DP
SCHEMATIC DIAGRAM OF
Intracellular (cytoplasmic)
In nucleated cells
Intracellular (cytoplasmic)
In APCs
Presentation of
Peptide-MHC I
to CD8+ CTL
PLASMA MEMBRANE
VIRAL
INFECTION
(viral Ag
production)
Transport of
Peptide-MHC I
Through Golgi
GOLGI
Step 1
Cytosolic Processing of
Endogenous Proteins
PROTEASOME
Peptide Driven
MHC-I Assembly
PEPTIDES
Step 3
TAP
ER
2-m
Step 2
Transporter in
Antigen Processing
Synthesis
MHC-I
Chainand
& -Microglobulin
Synthesis
of of
MHC-I
chain
2-microglobulin
Protein
Step 3
In Rough ER (RER)
TAP1
TAP2
57
Proteosome
A 700 kD form appears as a cylinder w/ stacked array
of 2 outer/ 2 inner rings composed of 7 subunits
3 of 7 subunits are catalytic sites for proteolysis
2 subunits LMP-2 and LMP-7 are encoded by genes
of the MHC
Performs basic housekeeping in cell- proteins
targeted for degradation are coated w/ ubiquitin
IFN gamma increases synthesis of LMP-2 and LMP-7
58
Presentation of
Peptide-MHC II to
CD4+ T-Helper Cell
Step 1 Endocytosis of
Plasma
membrane
Exogeneous Ag
into Endosom es
Exocytic vesicle
Binding of Peptide
to MHC-II
Endosome
Fusion of Endosome
with Vesicle Containing
MHC; Ii released.
Step 3
Processing of
Exogeneous Ag
in Endosom e
or Lysosom e
Lysosome
GOLGI
Step 2
MHC-I I Synthesis
Alpha
Beta
ER
Ii (invariant chain)
Step 1
2) Phagocytosis
3) Pinocytosis
Step 2
STEP 2: ENDOSOMAL
DEGRADATION OF
FOREIGN PROTEIN
1) Proteinases activated by acidity
2) Degraded to ~24 aa peptides
Steps 1-3
STEPS 1-3:
1) Receptor mediated exogenous
Ag uptake
2) Endosomal degradation of Ag
3) Loading peptide on MHC-II
STEP 3:
1) MHC-II interact with invariant
chain
2) CLIP production
3) HLA-DM assist peptide loading
on MHC-II
Step 3
4) HLA-DO block
HLA-DM activity
62
SUMMARY OF
Chloroquine
endocytosis
Emetine
Protein synthesis
64
Classical Presentation:
Endogenous antigen presentation by MHC-I to CD8+ T cells.
Exogenous antigen presentation by MHC-II to CD4+ T cells.
Non-classical Presentation:
Cross-presentation of exogenous antigens by MHC-I to CD8+ T
cells.
Non-peptide antigen presentation by CD1 to T cells and NK T
cells.
CROSS-PRESENTATION OF
1) Exogenous Ag uptake &
endosomal degradation
2) Peptide loading on MHC-I in RER?
3) Exogenous Ag peptide
presented in context of MHC-I
to CD8+ T cells.
EXOGENOUS
Ag BY MHC-I
CD1
H-2Kb
Line 2
Group 2
Line 5
Group 2
Group 1
68
69
70