Antigens and the Major

Histocompatibility Complex

CPMS
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Antigens vs. Immunogens
• Antigens = Substances that are able to combine to an antibody
• Immunogens = substances that cause immune response (eg,
release of antibodies)
• All immunogens are antigens; but not all antigens are
immunogens.
Factors that affectimmunogenicity
• Macromolecular size
• Most substances greater than 10,000 D induce immune response (there
are exceptions!). The general rule is, the bigger the molecule, the more it
induces an immune response
• Chemical Structure/Molecular complexity
• An antigen with a more complex structure would have the ability to elicit
immune response better than those with a simpler structure
• Foreignness
• An antigen derived from a taxonimically distant object elicit a better immune
response than antigens derived from a nearer taxa or the same taxa.
Factors that affectimmunogenicity
• Ability to be processed and presented by APCs
• They must be easily phagocytized, degraded, and presented by
MHC molecules to T cells
Factors influencing the immune response
• Age
• Overall Health
• Dose
• Route of inoculation
• Genetic
Nature of Epitopes
• Although an immunogen must have a molecular weight of at least
10,000, only a small part of the immunogen is actually recognized in
the immune response.
• This key portion of the immunogen is known as the determinant site
or epitope.
• Can be Linear (sequential) or Conformational
• B cells = they recognize unprocessed/unpresented epitopes
• T cells= they only recognize epitopes that are attach to MHC
molecules (Antigens that were presented to them)
Haptens
• Some substances are too small to be recognized by themselves, but
if they are complexed to larger molecules, they are then able to
stimulate a response. These substances are called HAPTENS.
• Haptens may be complexed artificially with a carrier or naturally
within the host
Classes of antigens
• Autoantigens
• Alloantigens
• Heteroantigens
• Heterophile
Adjuvants
• An adjuvant is a substance administered with an immunogen that
increases the immune response.
• It acts by producing a local inflammatory response that attracts a
large number of immune system cells to the injection sites
• Examples:
• Aluminum Salts (Only adjuvant approved in the US; used in HBV)
• Freund’s complete adjuvant
MAJOR HISTOCOMPATIBILITYCOMPLEX
MHC molecules
• Also called Human Leukocyte Antigens, they are molecules which
functions to bring antigen to the cell surface for recognition by T
cells, because T-cell activation will occur only when antigen is
combined with MHC molecules.

• They are encoded by a system of genes called the Major

Histocompatibility Complex.
Major HistocompatibilityComplex
• The major histocompatibility complex is a collection of genes
arrayed within a long continuous stretch of DNA on chromosome 6
in humans and on chromosome 17 in mice.

• The MHC is referred to as the HLA complex in humans and as the H-

2 complex in mice.

• Class I Molecules = Coded in A, B, and C Loci

• Class II Molecules = Coded in D region
• Class III Molecules = Coded in C4a, C4b and C2 regions
 MHC
• At each of these loci, or locations, there is the possibility of multiple
alleles. Alleles are alternate forms of a gene that code for slightly
different varieties of the same product.

• The MHC system is described as polymorphic, because there are so many

possible alleles at each location. For example, at least 580 different alleles
of HLA-A, 921 alleles of HLA-B, and 312 alleles of HLA-C have been
identified at this time.

• Uses of MHC = Disease correlation, Tissue compatibility, Paternity testing

Class I Molecules
• Found on all nucleated cells
• Highest on Lymphocytes, lower on liver cells, neural, muscle,
and sperm cells
• A glycoprotein dimer composed of alpha polypeptide chain
and β2-Microglobulin, where the alpha chain is furthermore
divided three domains, α1, α2, andα3
• HLA-A, HLA-B, and HLA-C
• Produced by ribosomes in the Endoplasmic reticulum
Class II molecules
• Found on Antigen Presenting Cells (B cells, Macrophages,
Dendritic cells, monocytes)
• Major class II molecules are: DR, DQ, and DP
• Composed of two of polypeptide chains, Alpha and Beta, both
with two domains; α1 and β1 is the peptide binding cleft
• Produced by Ribosomes in the Endoplasmic reticulum
Role of MHC molecules
• Remember that both Class I and Class II molecules functions to
perform antigen presentation to T cells

• T cells can only “see” and respond to antigens when they are
combined with MHC molecules.

• For an antigen to be properly presented:

• Peptides must be available in adequate supply
• They must bind MHC molecules effectively
• They must be recognized by the TCR
Role of Class Imolecules
• Class I molecules function to present endogenous peptides
(Viral proteins or intracellular products of tumor cells) to CD8
(Cytotoxic) cells
Class I Pathway
Role of Class IImolecules
• Class II molecules function to present exogenous peptides
(Bacterial antigens, or anything that can be endocytosed) to
CD4 (helper) cells
Class II Pathway
Clinical Significance of MHC
• Transplantation
• Autoimmune disorders
• Vaccine response and nonresponse
• Selective immunity
Disease associated in HLA
• HLA B-27: ankylosing spondylitis
• HLA DR3: associated with SLE
• HLA DR4: associated with RA
Methods for HLA Detection
• 1. Serological Method
• Test: Microlymphocytotoxicity testing/ Complement dependent cell cytotoxicity
• Test for both MHC Class I and II detection
• Inverted phase contrast microscope
• Usage of polyspecific AHG reagent
• ACD or phenol-free heparinized blood
• Purified T cell and/or B cell suspension for MHC Class I
• Purified B cell suspension for MHC Class II
• Preparations of B lymphocyte suspension:
• Nylon wool separation
• Fluorescent labeling
• Magnetic beads – method of choice
Methods for HLA Detection
• 2. Cellular method
• Test: Mixed lymphocyte reaction
• ACD or phenol-free heparinized blood
• For HLA-D related antigens
• Results are reported as counts per minute, stimulation index, or % relative response

• 3. Molecular method – preferred test for HLA antigens

• PCR (Polymerase chain reaction)
• Restricted Fragment length polymorphism (RFLP)