MAJOR HISTOCOMPATIBILITY COMPLEX
HLA/MHC
 Accounts for differences in how individuals respond to
particular immunogens
 HLA (Human Leukocyte Antigens)
 Dausset (French scientist)
 were first identified on circulating WBCs
 Aka MHC molecules because they determine whether
transplanted tissue is histocompatible and accepted, or
recognized as foreign and rejected
 MHC molecules
 Found not just on WBCs but on all nucleated cells in
the body
 Play a pivotal role in the development of both humoral
and cellular immunity
 Main function is to bring antigen to the cell surface for
recognition by T cells because only when antigen is
combined with MHC molecules does T cell activation
occur
 Clinically relevant because they may be involved in
transfusion reactions, graft rejection, and autoimmune
diseases
 Genes controlling expression of these molecules are the
system of genes known as the major
histocompatibility complex (MHC)
Genes Coding for MHC molecules (HLA Antigens)
MHC system
 Most polymorphic system found in humans
 Genes coding for the MHC molecules in humans are found
on the short arm of chromosome 6
 divided into 3 categories or classes
1. Class I –coded for at 3 different locations
or loci (A, B, and C)
2. Class II-genes are situated in the D region
- Different loci: DR, DQ, and DP
- There is a gene that codes for the alpha chain
and one or more genes that code for the beta chain
* Class I and II gene products are involved in antigen recognition
and influence repertoire of antigens to which T cells can respond
3. Class III
- Located on chromosome 6 situated between
the class I and class III regions
- Genes code for complement proteins and
cytokines (tumor necrosis factors alpha and
beta)
- Class III proteins are secreted proteins that
have an immune function, but they are not
expressed on cell surfaces
 Alleles
 Different forms of a gene that code for slightly
different varieties of the same product
 Examples: MHC (polymorphic)
HLA-A – 266 different alleles
HLA-B – 511 alleles
HLA-C – 128 alleles
 The probability that any two individuals will express
the same MHC molecules is very low
 An individual inherits two copies of chromosome 6,
and thus there is a possibility of two different alleles
for each gene on the chromosome, unless the
individual is homozygous (same alleles) at a given
location (genes are described as codominant)
 For the MHC system, each inherited chromosomal region
(haplotype) consists of a package of genes for A, B, C, DR,
DP, and DQ
 The full genotype would consist of two of each gene at a
particular allele
 Because there are numerous alleles or variant forms at each
locus, an individual’s MHC type is about as unique as a
fingerprint
 HLA Nomenclature
 Traditionally, HLA nomenclature has been defined
serologically through the use of a battery of antibodies
 Currently, advances in DNA analysis have made
identification of actual genes possible but the nomenclature
has become correspondingly more complex
 Example: HLA DRB1*1301
- Indicates the actual gene involved in coding for an
HLA DR1 antigen, with the B standing for the beta chain,
which is part of the antigen, and the 1301 indicating a
specific allele
 HLA antigens
 The uniqueness of the HLA antigens creates a major
problem in matching of organ donors because these
antigens are highly immunogenic
 However, in cases of disputed paternity,
polymorphisms can be used as a helpful identification
tool
 HLA testing can show that a man is not a
child’s father if the child has an HLA allele that
is not present in either the mother or the
alleged father or if the child does not possess
either of the two alleles that are present in the
alleged father at a given locus
 If the alleged father does in fact possess all the
alleles present in the child that are not from the
mother, he is likely a candidate, but paternity
cannot be absolutely proven
 Example:
Mother: HLA A2, A3 / B8, B12 / Cw1, Cw3
Child: HLA A3, A9 / B8, B16 / Cw2, Cw1
Alleged Father: HLA A1, A9 / B7, B14 / Cw2, Cw7
 Is the man the father of the child?
 Alleles not found in the mother are A9, B16, Cw2
 Father has A9 and Cw2, but not B16
 Result: The man is NOT the child’s father
 Structure of Class I Molecules
 Each of the MHC gene codes for a protein product
that appears on cell surfaces
 All of the proteins of a particular class share structural
similarities and are found on the same types of cells
 Class I MHC molecules are expressed on all
nucleated cells, although they differ in the level of
expression
MHC Class I Molecules
 Higher on lymphocytes
 Lowest on liver hepatocytes, neural cells, and muscle
cells (explains why HLA matching is not done in liver
transplant)
 HLA-C antigens are expressed at a lower level than
HLA-A and HLA-B antigens (2 most important to
match for transplantation)
 Each class I antigen is a glycoprotein dimer, made up
of two noncovalently linked polypeptide chains
 Alpha chain has as molecular weight of 45,000
 The lighter chain (ß2 – microglobulin)
associated with it has a molecular weight of
12,000 and is encoded by a single gene on
chromosome 15
 Folded into three domains: α1, α2, α3; is
inserted into the cell membrane via
hydrophobic regions
 The three external domains consist of about 90 amino
acids each
 The transmembrane domain has about 25
 hydrophobic amino acids along with a shorter stretch
of about 5 hydrophilic amino acids, and an anchor of
30 amino acids
 β2 – microglobulin does not penetrate the membrane
 α1 and α2 domains each form an alpha helix and that
these serve as the walls of a deep groove that function
as the peptide-binding site in antigen recognition
 Binding site is able to hold peptides that are
between 8 to 10 amino acids long
 Most of the polymorphism resides here
 α3 and β2 are similar to the constant regions found in
immunoglobulin molecules
 Nonclassical class I antigens
 Another group of molecules designated E, F, and G
 Are not expressed on cell surface except for G
 Do not function in antigen recognition but may play
other roles in the immune response
 G antigens are expressed on trophoblast cells during
the first trimester of pregnancy and are thought to
help ensure tolerance for the fetus
 Structure of Class II Molecules
 Occurrence is much more restricted because they are
found primarily on antigen presenting cells (B
lymphocytes, monocytes, macrophages, dendritic
cells, and endothelium
 Major class II molecules are DP, DQ, and DR
 Consist of two noncovalently bound
polypeptides that are both encoded by genes in
the MHC complex
 DR is expressed at the highest levels
 Both the α chain (mol.wt. 33,000) and the β chain
(mol.wt. 27,000) are anchored to the cell membrane
 Each has 2 domains
 Α1 and β1 domains come together to form the
peptide-binding site, similar to the one found in class
I molecules, however both ends of the binding cleft
are open, and this allows for capture of longer
peptides than is the case of class I molecules
 Nonclassical class II genes:
DM, DN, and DO
- Products of these genes play a
regulatory role in antigen processing
 The main role of class I and class II antigens is to
bind peptides within cells and transport them to the
plasma membrane where they can be recognized by T
cells
 Evolved to deal with 2 types of infectious agents
 Class I molecules present peptides that have been
synthesized within the cell to CD8 T cells
 Class II molecules contribute to antigen recognition
by CD4 T cells
 They mainly bind exogenous proteins, those taken
into the cell from the outside and degraded
 Class I molecules are the watchdogs of viral, tumor,
and certain parasitic antigens within the cell
 Class II molecules alert the CD4 T cells to the
presence of foreign proteins found outside the cell
 For a T cell response to be triggered
 Peptides must be available in adequate supply
for MHC molecules to bind
 They must be able to be bound effectively
 They must be recognized by the T cell receptor
 The difference in functioning of the two molecules is
tied to the mechanisms by which processed antigen is
transported to the surface
 Both types, however, must be capable of presenting
an enormous array of different antigenic
peptides to T cells
 The chemistry of the MHC antigens
controls what sorts of peptides fit in the
binding pockets
 Role of Class I Molecules
 Both class I and class II molecules are
synthesized in the rough ER
 Class I molecules bind peptides while still
in the ER
 Binding helps to stabilize the association of the
α chain with the β2- microglobulin
 Before binding with antigen occurs, newly
synthesized α chains freely bind calnexin
(keeps the α chain in a partially folded state
while it awaits binding to β2- microglobulin)
 When β2- microglobulin binds, calnexin is
released, and calreticulin and tapasin are
associated with the complex (help stabilize
complex for protein binding)
 Peptides that associate with the class I molecules are
approximately 9 AAs in length and derived from
partial proteolytic digestion of proteins previously
synthesized in the cytoplasm
 Digestion appears to be carried out by proteases in
large cylindrical cytoplasmic complexes
(proteosomes)
 Proteosomes
 Consist of approximately 16 to 20 components
that degrade proteins in the cytosol
 Once cleaved, the peptides are then pumped into the
lumen of ER in an ATP-dependent process by
specialized transporter proteins
 Transporters Associated with Antigen Processing
(TAP)
 Responsible for the ATP-dependent transport
from the cytoplasm to the lumen of ER of short
peptides
 TAP1 and TAP2 may function as “molecular
rulers” to measure the distance between the
amino and carboxyl termini of peptides so that
only peptides of the correct size are transported
 Most efficient at transporting peptides that are
12 residues or less in length
 Allelic difference in TAP proteins also influence the
immune response because they help determine the types
of peptides that are best transported
 Tapasin
 May help to bring TAP transporters into close
proximity to the newly formed MHC molecules so
that numerous peptides are available to them
 May also help in the loading of peptides into the class
 I molecules  On the cell surface, class II molecules are responsible for
 Once α chain has bound the peptide, the complex is rapidly formation of a trimolecular complex that occurs between
transported to the cell surface antigen, class II molecule, and an appropriate T cell receptor
 Of the thousands of peptides that may be processed in this  If binding occurs with a T cell receptor on a CD4+ T cell,
manner, only a small fractions of them (approximately 1 in the T helper cell recruits and triggers a B cell response,
2000) actually induce a response in association with class I resulting to antibody formation
molecules (Immunodominant)
 Immunodominant peptides Clinical Significance of MHC
 Peptides that bind with a strong affinity to class I  Testing for MHC antigens has typically been done
molecules and that produce enough complexes to find because:
CD8+ T cells with a complementary receptor  both class I and class II molecules are capable
 Binding is based on the interaction of only 2 or 3 of inducing a response that leads to graft
amino acid residues with the class I groove rejection
 Different class I molecules will have slightly different  Transplantation
binding affinities, and it is these small differences that  Play a role in development of autoimmune
determine to which particular antigen one individual will diseases
respond  Evidence that both class I and class II molecules play
 It is estimated that a single cell may express about 105 a major role in antigen presentation has more far-
copies of each class I molecules, so many different peptides reaching consequences
can be captured and expressed in this manner  They essentially determine the types of
 In healthy cells, self peptides are ignored by the T cells peptides to which an individual can mount an
 In diseased cells, peptides are derived from viral proteins or immune response (Ex. hepatitis B vaccine)
proteins associated with cancerous states
 Display of hundreds of class I molecules complexed to
antigen allows CD8+ T cells to continuously check cell
surfaces for the presence of other than self-antigen
 Role of Class II Molecules
 Class II molecules must be transported from the ER to
an endosomal compartment before they can bind
peptides
 While still in the ER, class II molecules associate with a
protein (Invariant chain), which may cause steric
obctruction and prevent interaction of the binding site
with any endogenous peptides in the ER
 Invariant chain
 A 31- kd protein that is made in excess so that enough
is available to bind with class II molecules shortly
after they are synthesized
 May be responsible for helping to bring α and β
chains together in the ER lumen then moving them
out through Golgi complex to the endocytic vesicles
 Also serves to protect the binding site of class II
molecules
 Once bound to the invariant chain, class II molecule is
transported to an endosomal compartment where it
encounters peptides derived from endocytosed, exogenous
proteins
 Antigen processing may help to unfold molecules and
uncover functional sites that are buried deep within the
native protein structure
 Invariant chain is degraded by a protease, leaving a small
fragment (corticotropin-like intermediate lobe peptide/
CLIP) attached to the peptide-binding cleft
 CLIP is then exchanged for exogenous peptides
 Selective binding of peptides may be promoted by:
 low pH of the endosomal compartment
 HLA-DM molecules help to mediate the reaction
 Generally, peptides of approximately 13 to 18 amino acid
residues can bind
 Conserved amino acids are distributed all along the actual
binding site
 Allows hydrogen binding to take place along the
length of the captured peptide
 There are also several pockets in the class II proteins that
easily accommodate amino acid side chains
 This gives class II proteins more flexibility in the
types of peptides that can be bound
 Once binding has occurred, class II protein-peptide complex
is transported to the cell surface