Transcript 3-ag-t cells - PowerPoint Presentation

Lectures 5 & 6
Antigen Recognition
by T Lymphocytes
Lecture objectives
•
Big questions?
How TCR recognizes antigens?
Why many autoimmune diseases are associated with particular types of MHC genotypes?
Why MHC molecules are the major antigens responsible for transplantation rejections?
•
•
•
•
•
•
•
•
•
•
•
•
•
•
Structure of TCRs
CDRs
Differences of recognized antigen between TCR and immunoglobulins (Ig)
TCR germ line configuration and rearrangements
TCR specificity
Clonality of T cells
Differences between TCR a and b chains
Names of human MHC I and II genes.
How antigens are processed for presentation on MHC I and II?
MHC class I and II molecules sample and present antigens of different origins. How?
MHC polymorphism
How many different MHCs a person can express? Why?
Structures of MHC I and II
HLA typing
Core content
A big picture:
How do T cells recognize antigens?
MHC molecule
TCR
Similarity between TCR and Ig
Both:
• Bind antigen
• Have variable region
• Constant region
• Each binding site is a
heterodimer
(composed of 2
different chains)
TCRs act only as
receptors
Igs act as receptors and
effector molecules
(soluble antigenbinding molecules)
Figure 3-2
TCR genes undergo DNA rearrangement in thymus
*No Ds in Va gene; DJ first then VDJ in b gene rearrangement
Un-rearranged
rearranged
expression
rearranged
Un-rearranged
If you can not make antigen receptors as in the case of SCID patients,
Figure 3-4
you can not make T and B cells, and are susceptible even to
opportunistic pathogens (e.g. C. albicans)
TCR complex
Figure 3-6
CD3 chains transmit signals
Figure 3-7
Figure 3-8 part 1 of 2
Figure 3-8 part 2 of 2
gd T cells recognize a limited set of antigens:
•Host MHC class 1b: T10/22, MICA/B
•Nonproetin alkylamines
•Bacterial products: Mycobacterial Hsp, superantigens
(SEA)
•Host heat shock proteins
•Transformed or stressed host cells
Antigen processing is required to present antigen peptides to TCR.
TCRs bind short antigen peptides but not whole antigen proteins
Figure 3-9
The structures of CD4 and CD8
Figure 3-10
MHC class I molecules present antigens to CD8+ T cells, and
MHC class II molecules present antigens to CD4+ T cells:
Figure 3-12
Figure 3-13 part 1 of 2
Figure 3-13 part 2 of 2
Figure 3-14
The peptide-binding groove of MHC molecules
Figure 3-15
Figure 3-16
Two different types of antigens:
Extracellular for MHC II and intracellular for MHC I
They are generated in different compartments
Transport of Cytosolic Peptides into ER
Role of transporter associated with antigen processing, TAP
Figure 3-17
Bare lymphocyte syndrome: No TAP
expression  no MHC I expression 
no CD8 T cells  defective cytotoxic
activity against virus-infected cells
chronic respiratory infection
TAP=TAP1+TAP2
Proteosome: a barrel shaped protein
complex composed of 28 subunits
8 or more AA-long polypeptides are
transported
MHC I cannot leave ER without loaded
peptides
Assembly of antigen peptide/MHC class I complex.
Molecular chaperons (calnexin, calreticulin and tapasin)
aid the folding of MHC I and loading of peptides
Figure 3-18
In the absence of infection, self peptides are presented on MHC, but do not activate T cells.
A HSV protein inhibits TAP function, and an Adeno virus protein inhibits MHC I expression.
Assembly of antigen peptide/MHC class II complex
-Extracellular microorganisms are taken up by macrophages via phagocytosis, and by B
cells by cell surface Ig-mediated endocytosis
-MHC II molecules bind peptides in the fused vesicles, not in ER
-Invariant chain, CLIP and HLA-DM guide the peptide loading
-After losing CLIP, MHC II must bind peptides or gets degraded.
-Certain pathogens (e.g. mycobacteria), when engulfed, prevent the fusion of
phagosomes and lysosomes, and persist in phagosomes.
Figure 3-20
Figure 3-21
TCR recognition of antigens induce T cell activation,
functional maturation, and killing/activation of target cells
Figure 3-11
Different cell distribution of MHC class I and II
Figure 3-22
• All cells express MHC I for
comprehensive surveillance by
CD8 T cells
• Only some cells express high
levels of MHC II and MHC I
• These are B cells, macrophages,
dendritic cells and thymic
epithelial cells.
• B cells, macrophages and
dendritic cells are called
professional antigen- presenting
cells (APC).
• IFN-g increases the expression
of MHC II in APC and induces
the expression in non-APC cells
at sites of infection
Figure 3-23
Figure 3-24 part 1 of 2
CD8/NK
NK
Remains intracellular
NK
MHC CLASS I molecules form ligands to activate CD8+ cells
and inhibit NK cells
Polymorphism: presence of multiple
alternative forms (alleles) of a gene.
Help peptide loading
With antigen peptides
activate CD4+ T cells
Polymorphism allows the population can handle a variety of pathogens.
Major Histocompatibility Complex
(=HLA locus)
• MHC molecules in human is also called HLA (human leukocyte antigen)
Class I and II locus.
• HLA-DR alpha chain is monomorphic
• HLA-DRB1 is most polymorphic in MHC II genes
• HLA-DRB1 is always present in any individual
• HLA-DRB3/4/5 is present in some but not all people.
• A heterozygote person expresses two haplotypes.
• A person can express 3-6 class I and 3-8 class II isoforms.
• 2198 possible class II isoforms in the human population.
• 440 MHC I isoforms in the human population.
• [MHC isoforms]  [presentable antigen peptides]  [TCR repertoire]
Figure 3-25
Figure 3-27
Genetics of MHC gene expression:
both alleles are expressed (co-dominant)
• In any mating, four possible
combinations of haplotypes can
be found in the offspring; thus
siblings are also likely to differ in
the MHC allele they express.
• Halplotype: The particular
combination of MHC alleles
found on a given chromosom 6.
Figure 3-26
Figure 3-28
Each MHC isoform binds a characteristic set of peptides
-Anchor residues in peptides are important for binding to MHC
-Not all residues are important
Figure 3-29
Degenerate binding allows each MHC molecule handles many peptides.
T cell receptor recognition of antigens is MHC-restricted
Figure 3-30
Being heterozygous for MHC is
advantageous in presenting antigens
Figure 3-31
Pathogens can select MHC polymorphism in human population
A scenario:
Selection 1 as the
result of multiple
successive infections
Figure 3-32
Selection 2 as
the result of
epidemic of a
new pathogen
MHC Heterozygocity makes AIDS+ people more resistant to HIV
Complete Heterozygocity
Homozygocity
HLA typing: how?
Microcytotoxicity assay for detection of HLA antigens
Anti-HLA serum, or monoclonal antibody, is mixed with live lymphocytes.
Specific Ig binds to the polymorphic protein moiety of the HLA molecule
expressed on the cell surface. Exogenous complement is added to the well
which will result in lysis of cells to which antibody has been bound. Cell death
is determined by ethidium bromide vital stain exclusion.
Flow cytometry or ELISA
Monoclonal antibodies to different MHC alleles have been generated.
Using panels of these antibodies, HLA typing before transplantation is
possible.
RFLP: Restriction Fragment Length Polymorphism
Digestion of genomic DNA with certain restriction enzymes followed by
hybridization with radio-labeled MHC gene probes gives MHC isotypespecific digestion patterns.
PCR: Polymerase Chain Reaction
PCR using MHC gene-specific primers