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MICROBIOLOGY AND IMMUNOLOGY MOBILE - IMMUNOLOGY CHAPTER TEN
MAJOR HISTOCOMPATIBILITY COMPLEX (MHC)
AND T-CELL RECEPTORS - ROLE IN IMMUNE RESPONSES
I. Historical Overview
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). Studies with inbred strains of mice showed that genes in the MHC were also involved in controlling both humoral and cell-mediated immune responses. For example, some strains of mice could respond to a particular antigen but other strains could not and these strains differed only in one or more of the genes in the MHC. Subsequent studies showed that there were two kinds of molecules encoded by the MHC – Class I molecules and class II molecules. Class I molecules were found on all nucleated cells (not red blood cells) whereas class II molecules were found only on antigen presenting cells, (APCs) which included dendritic cells, macrophages, B cells and a few other types (Figure 1).
It was not until the discovery of how the T cell receptor (TCR) recognizes
antigen that the role of MHC genes in immune responses was understood. The TCR
was shown to recognize antigenic peptides in association with MHC molecules. T
cells recognize portions of protein antigens that are bound non-covalently to
MHC gene products. Cytotoxic T cells (Tc) recognize peptides bound to class I
MHC molecules and helper T cells (Th) recognize peptides bound to class II MHC
molecules. The three dimensional structures of MHC molecules and the TCR have
been determined by X-ray crystallography so that a clear picture of how the TCR,
MHC gene products and antigen interact has emerged.
II. Structure of 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 phosphoylation and binding to cytoskeletal elements. Second, a transmembrane region containing hydrophic amino acids by which the molecule is anchored in the cell membrane. Third, a highly conserved α3 immunoglubilin-like domain to which CD8 binds. Fourth, a highly polymorphic peptide binding region formed from the α1 and α2 domains. The β2- microglobulin associates with the α chain and helps maintain the proper conformation of the molecule.
An analysis of which part of the class I MHC molecules is most variable demonstrates that variability is most pronounced in the α1 and α2 domains, which comprise the peptide binding region (Figure 3). The structure of the peptide binding groove, revealed by X-ray crystallography, shows that the groove is composed of two α helices forming a wall on each side and eight β-pleated sheets forming a floor. The peptide is bound in the groove and the residues that line the groove make contact with the peptide (Figure 4). These are the residues that are the most polymorphic. The groove will accommodate peptides of approximately 8-10 amino acids long. Whether a particular peptide will bind to the groove will depend on the amino acids that line the groove. Because class I molecules are polymorphic, different class I molecules will bind different peptides. Each class I molecule will bind only certain peptides and will have a set of criteria that a peptide must have in order to bind to the groove. For example, Figure 5 shows that one class I molecule will bind peptides that have a leucine (L) as the carboxy-terminal amino acid and either tyrosine (Y) or phenylalanine (F) as the 4th amino acid from the carboxy-terminal end. As long as these two conditions are met a peptide will bind, regardless of what the other amino acids are. Similarly a different class I molecule will bind any peptide that has a tyrosine (Y) as the second amino acid from the amino-terminal end and either a valine (V), isoleucine (I) or leucine (L) at the carboxy-terminal end (Figure 5). Thus, for every class I molecule, there are certain amino acids that must be a particular location in the peptide before it will bind to the MHC molecule. These sites in the peptide are referred to as the “anchor sites”.
Within the MHC there are 6 genes that encode class I molecules HLA-A, HLA –B,
HLA-C, HLA-E, HLA-F and HLA-G. Among these HLA-A, HLA –B, and HLA-C are the most
important and are most polymorphic. Table 1 shows the degree of polymorphism at
each of these loci.
Table 1. Polymorphism of class I MHC genes | |
Locus |
Number
of alleles (allotypes) |
HLA-A | 218 |
HLA-B | 439 |
HLA-C | 96 |
HLA-E, HLA-F and HLA-G | Relatively few alleles |
III. Structure of 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 phosphoylation and binding to
cytoskeletal elements; second, a transmembrane region containing hydrophic amino
acids by which the molecule is anchored in the cell membrane, third, a highly
conserved α2 domain and a highly conserved β2 domain to which CD4 binds and
fourth, a highly polymorphic peptide binding region formed from the α1 and β1
domains.
As with Class I MHC molecules, an analysis of which part of the class II MHC
molecule is most variable demonstrates that variability is most pronounced in
the α1 and β1 domains, which comprise the peptide binding region (Figure 7). The
structure of the peptide binding groove, revealed by X-ray crystallography,
shows that, like class I MHC molecules, the groove is composed of two α helices
forming a wall on each side and eight β-pleated sheets forming a floor. Both the
α1 and β1 chain contribute to the peptide binding groove. The peptide is bound
in the groove and the residues that line the groove make contact with the
peptide. These are the residues that are the most polymorphic. The groove of
Class II molecules is open at one end so that the groove can accommodate longer
peptides of approximately 13-25 amino acids long with some of the amino acids
located outside of the groove. Whether a particular peptide will bind to the
groove will depend on the amino acids that line the groove. Because class II
molecules are polymorphic, different class II molecules will bind different
peptides. Like class I molecules, each class II molecule will bind only certain
peptides and will have a set of criteria that a peptide must have in order to
bind to the groove (i.e. “anchor sites”).
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. Table 2
shows the degree of polymorphism at each of these loci.
IV. Important Aspects of MHC
A. Although there is a high degree of polymorphism for a species, an individual has maximum of six different class I MHC products and only slightly more class II MHC products (considering only the major loci).
B. Each MHC molecule has only one binding site. The different peptides a given MHC molecule can bind all bind to the same site, but only one at a time.
C. Because each MHC molecule can bind many different peptides, binding is termed degenerate.
D. MHC polymorphism is determined only in the germline. There are no recombinational mechanisms for generating diversity.
E. MHC molecules are membrane-bound; recognition by T cells requires cell-cell contact.
F. Alleles for MHC genes are co-dominant. Each MHC gene product is expressed on the cell surface of an individual nucleated cell.
G. A peptide must associate with a given MHC of that individual, otherwise no immune response can occur. That is one level of control.
H. Mature T cells must have a T cell receptor that recognizes the peptide associated with MHC. This is the second level of control.
I. Cytokines (especially interferon-γ) increase level of expression of MHC.
J. Peptides from the cytosol associate with class I MHC and are recognized by Tc cells. Peptides from within vesicles associate with class II MHC and are recognized by Th cells.
K. Polymorphism in MHC is important for survival of the species.
Table 2. Polymorphism of class II MHC genes | |
Locus |
Number
of alleles (allotypes) |
HLA-DPA HLA-DPB |
12 88 |
HLA-DQA HLA-DQB |
17 42 |
HLA-DRA HLA-DRB1 HLA-DRB3 HLA-DRB4 HLA-DRB5 |
2 269 30 7 12 |
HLA-DM and HLA-DO | Relatively few alleles |
V. Structure of the T cell receptor (TCR)
The TCR is a heterodimer composed of one α and one β chain of approximately
equal length (Figure 8). Each chain has a short cytoplasmic tail but it is to
small to be able to transduce an activation signal to the cell. Both chains have
a transmembrane region comprised of hydrophobic amino acids by which the
molecule is anchored in the cell membrane. Both chains have a constant region
and a variable region similar to the immunoglobulin chains. The variable region
of both chains contains hypervariable regions that determine the specificity for
antigen. Each T cell bears a TCR of only one specificity (i.e. there is allelic
exclusion).
The genetic basis for the generation of the vast array of antigen receptors on B
cells has been discussed previously (see lecture on Ig genetics). The generation
of a vast array of TCRs is accomplished by similar mechanism. The germline genes
for the TCR β genes are composed of V, D and J gene segments that rearrange
during T cell development to produce many different TCR β chains (Figure 9). The
germline genes for the TCR α genes are composed of V and J gene segments which
rearrange to produce α chains. The specificity of the TCR is determined by the
combination of α and β chains.
There is a small population of T cells that express TCRs that have γ and δ
chains instead of α and β chains. These gamma/delta T cells predominate in the
mucosal epithelium and have a repertoire biased toward certain bacterial and
viral antigens. The genes for the δ chains have V, D and J gene segments whereas
the genes for the γ chains have only V and J gene segments but the repertoire is
considerably smaller that than that of the alpha/beta T cells. The gamma/delta T
cells recognize antigen in an MHC-independent manner unlike the alpha/beta T
cells.
TABLE 3 |
||
GENES |
||
Properties |
Ig |
TCR |
Many VDJs, Few C's |
Yes |
Yes |
VDJ Rearrangement |
Yes |
Yes |
V pairs form antigen-recognition site |
Yes |
Yes |
Somatic hypermutation |
Yes |
No |
PROTEINS |
||
Transmembrane forms |
Yes |
Yes |
Secreted forms |
Yes |
No |
Isotypes with distinct functions |
Yes |
No |
Valency |
2 |
1 |
Adapted from Janeway and Travers, Immunobiology |
VI. TCR and CD3 Complex
The TCR is closely associated with a group of 5 proteins collectively called
the CD3 complex (Figure 10). The CD3 complex is composed of one γ, one δ, two ε
and 2 ξ chains. All of the proteins of the CD3 complex are invariant and they do
not contribute to the specificity in any way. The CD 3 complex is necessary for
cell surface expression of the TCR during T cell development. In addition, the
CD3 complex transduces activation signals to the cell following antigen
interaction with the TCR.
VII. The “Immunological synapse”
The interaction between the TCR and MHC molecules are not very strong. Accessory
molecules are necessary to help stabilize the interaction (Figure 11a,b). These
include: 1) CD4 binding to Class II MCH, which ensures that Th cells only
interact with APCs; 2) CD8 binding to class I MHC, which ensures that Tc cells
can interact with target cells; 3) CD2binding to LFA-3 and 4) LFA-1 binding to
ICAM-1. The accessory molecules are invariant and do not contribute to the
specificity of the interaction, which is solely determined by the TCR. The
expression of accessory molecules can be increased in response to cytokine,
which is one way that cytokines can modulate immune responses.
In addition to accessory molecules which help stabilize the interaction between
the TCR and antigen in association with MHC molecules, other molecules are also
needed for T cell activation. Two signals are required for T cell activation –
one is the engagement of the TCR with Ag/MHC and the other signal comes from the
engagement of co-stimulatory molecules with their ligands. One of the most
important (but not the only) co-stimulatory molecule is CD28 on T cells which
must interact with B7-1 (CD80) or B7-2 (CD81) on APCs . Like accessory molecules
the co-stimulatory molecules are invariant and do not contribute to the
specificity of the interaction. The multiple interactions of TCR with Ag/MHC and
the accessory and co-stimulatory molecules with their ligands have been termed
the “immunological synapse.”
Not only is co-stimulation necessary for T cell activation, a lack of
co-stimulation may result in anergy (i.e., inability to respond to antigen) or
down-regulation of the response. Figure 12 shows the possible outcomes of a T
cell receiving one or both of the signals necessary for activation. Engagement
of the TCR with Ag/MHC but no co-simulation results in anergy. Engagement of
only the co-stimulatory molecule has no effect. Engagement of TCR with Ag/MHC
and co-stimulatory molecules with their ligand results in activation. Engagement
of the TCR with Ag/MHC and engagement of B7 ligand with CTLA-4, molecules
similar to CD28, results in down-regulation of the response. CTLA-4/B7
interaction sends an inhibitory signal to the T cell rather than an activating
signal. This is one of the ways that immune responses are regulated. CTLA-4 is
expressed on T cells later in an immune response and this helps to turn off the
response.
VIII. Key Steps in T cell Activation
A. APC must process and present peptides to T cells
B. T cells must receive a co-stimulatory signal - usually from CD28/B7
C. Accessory adhesion molecules must help to stabilize the binding of T cells and APC. (CD4/class II MHC, CD8/classs I MHC, LFA-1/ICAM-1 and CD2/LFA-3)
D. Signals from cell surface must be transmitted to the nucleus via second messengers
E. Cytokines, including IL-2, must help drive cell division
TABLE 4 |
|
T cell molecule |
Ligand on second cell |
CD4 on helper T cells | class II MHC molecules |
CD8 on cytotoxic T cells | class I MHC molecules |
LFA-2 (CD2) | LFA-3 |
LFA-1 | ICAM-1, ICAM-2 |
LFA = Leukocyte Function-associated Antigen | |
ICAM = Intercellular Adhesion Molecule |
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