Adaptive immunity - T-Cell receptors & MHC proteins Flashcards
What are the 2 types of T-cells /T-cell receptors and how are they defined
- only expressed on membranes, not as soluble proteins
- T helper cells (CD4 +ve)
- Augment immune responses
- T cytotoxic cells (CD8 +ve)
- Specifically kill infected host cells
- act a bit like NK cells but much more specific
- only kill host cells infected with a particular pathogen that they can recognise with their specific T-cell receptors
- Receptor structure on both subpopulations is the same!
Describe the basic structure of the T cell receptor
- Broadly fab-like structure
- TCR consists of an 𝛼 and 𝛽 chain
- Each chain has a Variable and Constant region
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Extracellular domains of the T cell receptor are homologous to the variable and constant regions of immunoglobulins.
- each V region contains 3 CDRs (hyper variable)
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Extracellular domains of the T cell receptor are homologous to the variable and constant regions of immunoglobulins.
- Stalk segments under constant regions connected by disulfide bonds
- Transmembrane region
- Cytoplasmic tail
- As the TCR is at the membranes of T cells, it has hydrophobic aa residues on c-terminus of the chains
- A subset of T cells (1-5%) express 𝛾 𝜹 (gamma delta) receptors (chains) instead of 𝛼 𝛽
- less diverse
- V𝛼 and V𝛽 domains each have 3 CDRs (1-3)
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CDR3 regions of 𝛼 and 𝛽 chains are the most variable
- same as for antibodies:
- CDR3 loop region is the most variable
- same as for antibodies:
Can the T- receptor signal by itself?
explain your answer
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Expression of the T-cell receptor on the cell surface requires association with additional proteins.
- when it recognises antigen it has to associate with additional proteins for signalling to occur. it can’t signal by itsself
- these proteins are known collectively as CD3
- this gives rise to the T-cell receptor Complex containing:
- The T cell receptor complex:
- 𝛼 and β subunits (TCR)
- which have to associate with →
- CD3 subunits (ε, δ and γ) and ζ
- E + D, E+Y and 2 Zeta stalks
- 𝛼 and β subunits (TCR)
- Required for optimal cell surface expression and signalling
- CD3 subunits contain ITAMs (Immunoreceptor Tyrosine Activation Motifs) in their cytoplasmic regions
- also found in alpha and beta chains that interact with B cell receptors
- antigen binding → ITAMs phosphorylated → downstream signalling
- when it recognises antigen it has to associate with additional proteins for signalling to occur. it can’t signal by itsself
Draw a diagram of the T-cell receptor complex
see lecture notes: 4th tog
Describe the TCR genes
- Similar to antibody receptor genes: 2 genes loci for alpha and beta chains
- Chromosome 14:
- contains alpha 2 exons,
- V region and many J regions and c domains
- a bit like the light chain of antibodies
- Chromosome 7
- V segments, J segments and D segments and c domains
Does somatic V(D)J recombination occur in T cells? if it does, explain its similarities
exact same as B cell gene rearrangement
only difference is that it occurs in the thymus and not the bone marrow
Describe the diversity of T cell receptor genes
T cells generally have more gene segments than B cells, creates bigger junctional diversity compared to B cells.
- Multiple copies of V region gene segment [Vn x Jn/Vn x Dn x Jn]
V𝛼 ~70 segments, J𝛼 61, V𝛽 = 52, D𝛽 2, J𝛽 13
whereas for B cell its: max 6, 23, 40 - 𝛼 x 𝛽 chain combination [Va x Ja] x [Vb x Db x Jb] =~6*10^6
- Junctional diversity = ~2*10^11
- Concentrated in the CDR3s of TCR 𝛼 and 𝛽 chains
- i.e. these are the most variable
- CDR1 and 2 are encoded in germline
- CDR3 is the VJ/VDJ join so is very diverse
- Total diversity = ~10^18
- Concentrated in the CDR3s of TCR 𝛼 and 𝛽 chains
Compare the differences in diversity of B and T cell repertoire
B-cells undergo somatic hypermutation which T-cells don’t, however:
The T-cell repertoire is more diverse because T cells generally have more gene segments than B cells -> creates bigger junctional diversity compared to B cells
T-cell repertoire: 10^18
B-cell repertoire: 10^14
What is the main big difference between B-cell antibodies and TCRs?
The V regions of TCRs do NOT undergo somatic mutation!!
Why do V regions of TCRs not undergo somatic hypermutation
- why?
- maybe its too dangerous
- if T cells somaticly mutate, you might get receptors recognising own tissues
- B cells the same may occur but B cells need T cells
- or they may not just need that high affinity generation
- real answer: we don’t know for sure
- maybe its too dangerous
Do B and T cells recognise the same antigen? what type of pathogens are they important for dealing with
- B and T cells recognise different types of antigens
- B cell immunity is particularly important in defence against extracellular pathogens
- generally true
- can bind to structures on surface of pathogens or surface of viruses etc.
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B cells recognise free, “native” antigens
- i.e. non-cell associated antigen (unprocessed antigens)
T cells don’t recognise free, “native” antigens
How do T cells recognise antigen?
- T cells also important in dealing with intracellular pathogens
- How can T cells recognise intracellular antigens? TCR can’t look inside the cell so how?
- samples of whatever is inside a cell is displayed on the surface so a T cell wit hthe right receptor can recognise wether or not a cell is infected. so how do they do this?
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Major Histocompatibility proteins (MHC)
- Display bits of whatever is inside the cell on the surface.
- If infected, T cell will recognise the non-self presentation i.e. antigen
- So T cells don’t recognise native antigens, only processed antigens!!
- antigen must be processed (degraded into smaller peptides)
- antigen binds to MHC and MHC brings it to the cell surface and presents it
- Protein → peptide → MHC → cell surface → T cell recognition
- T cells recognise “cell-associated, processed antigen
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NB! B cells antibodies react to naive unprocessed antigen, whilst T cells and their TCRs recognise “cell-associated” processed antigen!!!
- T cells require antigen to be presented to them
Describe the genes and nature of said genes that encode MHC and their importance
- T cells require antigen presentation by cells expressing Major Histocompatibility proteins (MHC)
- Discovered during research on graft rejection. Encoded by the genes of the Major Histocompatibility Complex Chromosome 6 (in humans)
- also known as HLA molecules in humans (human leucocyte antigen). the genes!! not the protein
- **e.g. HLA-A, HLA-B, HLA-C
- 3 gene loci coding for 3 different MHC proteins!
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very polymorphic
- e.g. > 1400 alleles of HLA-B locus in the human population. Alleles may differ by up to 20 a.a. substitutions
- so when you match for transplant, you try to match these genes as close together as possible to avoid graft rejection
- major role in antigen presentation and initiation of T cell responses
Describe the concept of MHC restriction
- T lymphocytes can only recognise antigen in the context of self-MHC molecules
- this is known as MHC restriction
- Experiments with inbred mouse strains and virally infected cells (had the same MHC proteins on their surfaces)
- Mouse Strain A and mouse strain B immunised with Virus, T cell from the mice were isolated and cultured in vitro with cells infected with the same virus
- if you took T cells from mouse strain A and mixed with cells from mouse A → would kill mouse A infected cells
- if you took T cells from mouse strain B and mixed with cells from Mouse B → T cells can’t kill infected cells from mouse A
- SO T cells will only recognise antigen thats being presented to them by ?? (31 min)
- why did this happen? → 2 ideas
- 2 receptors on T cells – one (TCR) for antigen, one for MHC?
- 1 thats recognised antigen, and one recognising MHC
- 1 receptor on T cells (TCR) – recognises antigen + MHC?
- only 1 receptor on t cell that recognises antigen and MHC
- 2 receptors on T cells – one (TCR) for antigen, one for MHC?
x-ray crystallography proved answer
- Unknown peptide antigen bound as part of the structure
- At the tip of the molecule in a groove, there was a peptide → proved that T cells recognise MHC and foreign peptide.
- MHC, they bind foreign peptide and transport it to the cell surface
Describe the structure of a TCR binding MHC1+peptide
- FAB with CDRs in the middle, MHC protein on the bottom. Peptide in yellow
- Crystallographic studies demonstrated:
- **(1) MHC binds peptide
- (2) TCR recognises complex of peptide + self-MHC
- The loops
- CDR1 and CDR2 of T cell receptor bind self MHC
- CDR1 and CDR2 inherited in germline so less variable
- CDR3 binds to the peptide
- hypervariable
- CDR1 and CDR2 of T cell receptor bind self MHC
- Another explanation idea for why somatic mutation doesnt occur in T cells: This may that if T cell receptor genes mutated → you would loose recognition of self MHC if these CDRs mutate
- CDR1 and CDR2 bind self MHC (germline-encoded)
- CDR3 binds peptide (variation introduced by junctional diversity)
Describe the 2 classes of MHC and their roles and what antigen presenting cells are
- Two classes of MHC proteins are involved in antigen recognition by T cells
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MHC class I
- Expressed by all nucleated cells
- so basically all cells apart from red blood cells
- Present peptides derived from endogenous proteins to cytotoxic (CD8) T cells
- endogenous meaning: has to be synthesised by the cell that will display it
- Expressed by all nucleated cells
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MHC class II
- Expressed by certain leucocytes (dendritic cells, B cells, macrophages).
- more restricted expression pattern
- Present peptides derived from exogenous proteins to helper (CD4) T cells
- things taken up by a cell (exogenous)
- Expressed by certain leucocytes (dendritic cells, B cells, macrophages).
- Antigen presenting cells: Dendritic cells, B cells and macrophages
- so only the ones that present to T helper cells
Describe the protein structure of MHC1 and MHC2
- quite similar
- MHC1:
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Polymorphic transmembrane alpha chain, invariant ß-microglobulin
- invariant = just stabilises molecule doesn’t change
- alpha chain intrinsic, beta microglobulin just stbilises
- only alpha is transmembrane
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Polymorphic transmembrane alpha chain, invariant ß-microglobulin
- MHC2:
- Polymorphic transmembrane alpha and beta chains
- both transmembrane
- domains further from the membrane → more polymorphic and bind to peptide
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Membrane -proximal domains are Ig-like
- the ones close to membrane
- Membrane distal domains bind peptide
- Membrane distal domains contain polymorphisms
Describe how MHC 1 and MHC 2 bind peptides
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MHCI bind peptides 8-10 a.a. long
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N and C-termini of peptides bind to invariant sites at ends of the groove.
- invariant residues present at the ends of the grooves
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Two or three “anchor residues” on the peptides bind to “specificity pockets” formed by polymorphic residues.
- at the base of the groove
- fairly closed (restricts length of peptide that can bind)
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N and C-termini of peptides bind to invariant sites at ends of the groove.
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MHCII bind peptides ~ 13-18 a.a. long
- Peptide backbone interacts with conserved residues that line the base of groove.
- “Anchor residues” on the peptide bind to “specificity pockets” formed by polymorphic residues at the end of the grooves
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MHCI bind peptides 8-10 a.a. long
Does MHC 1 bind bigger or smaller peptide? what’s a good analogy of MHC peptide binding
- MHC1 smaller peptide
- MHC II bigger peptide
- Hotdog analogy:
- MHC1 is the bun, peptide is the sausage
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A particular MHC molecule with a particular sequence (allele) can bind a wide range of related peptides (structurally related)
- the diversity of the MHC proteins is inhertited, no somatic recombination etc.
- MHC1 smaller peptide
- MHC II bigger peptide
- Hotdog analogy:
- MHC1 is the bun, peptide is the sausage
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A particular MHC molecule with a particular sequence (allele) can bind a wide range of related peptides (structurally related)
- the diversity of the MHC proteins is inhertited, no somatic recombination etc.
- looking from above down onto the molecule
- beta sheets, and alpha helical region coming out of the surface formign the sides of the domain
Describe the process of antigen presentation by MHC1
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Antigen presentation by MHCI (endogenous antigen)e.g. virus-infected cell to cytotoxic T cell (CD8 +ve)protein made from within the cell (endogenous)
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Proteosome:
- multi-subunit complex that breaks down misfolded proteins
- present in all cells
- i.e. will try to break down viral proteins too
- another subunit will join the proteosome creating a → immunoproteosome
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Immunoproteosome:
- formed when another subunit is produced induced by interferon
- peptides transported to ER by ATP-hydrolysis driven transporter, TAP (transporter associated with antigen presentation).
- in ER, antigen can bind MHC1
- MHC1 protein will be destined to be sent up to the cell surface regardless of antigen binding or not
- peptides loaded onto MHCI in ER
- MHCI-peptide transported to cell surface for recognition by cytotoxic T cell
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Proteosome:
Describe the process of antigen presentation by MHC2
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Antigen presentation by MHCII (exogenous antigen)
- cell isn’t infected! but needs help from a helper T cell to get the immune response going
- antigen taken up by phagocytosis or endocytosis → in phagolysosome (if phagocyte) enzymes break down protein of pathogen into peptides → peptides associate with MHC II in the endocytic compartments (vacuoles) (in MHC1 association happens in the ER)→ MHC II take peptides to surface where they can be recognised by T helper cell with the right receptor
- antigen taken up by phagocytosis or endocytosis
- acidification in vesicles promotes unfolding and proteolysis
- peptides associate with MHC II in the endocytic compartment
- MHCII-peptide transported to cell surface for recognition by helper T cell
Describe cross-presentation by MHC 1 and MHC2
- another type of antigen presentation called cross-presentation
- some circumstances wehre a dendritic cell needs to induce a naive cytotoxic T cell to respond to antigen. The D cell is not itsself infected, infection in other tissue maybe etc, none the less they need to induce the naive T cytotoxic cell to respond
- not completely understood
- Exogenous peptide can associate with MHC1
- shown below: necrotic cell (aren’t infected just dying), dendritic cell may take up a necrotic cell and in endosomal or in the cytosol the antigen can associate with MHC1
- this is important because it allows anitgen to be presented to Cytotoxic T cells even if the cell itsself isn’t infected → this might be important in the responses Cytotoxic T cells make to some tumours
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CROSS-PRESENTATION
- Some dendritic cells present exogenous peptide associated with MHCI to cytotoxic T cells
- Allows antigen presentation to cytotoxic T cells without the dendritic cells themselves being infected
- Important in cytotoxic T cell responses to many tumours
What other roles do MHC proteins have
T cell signalling
Thymic selection
Describe the role of MHC proteins in T cell signalling
- also play a role in activation of T cells.
- For T cell activation, in addition to the TCR complex (αβ chains +CD3-proteins (to recognise antigen) + 𝜁 chain), co-receptors are also required to
- 1) stabilise the interaction
- 2) facilitate signalling
- in MHC2 antigen presentation is CD4, in MHC2 antigen presentation the co-receptor is CD8
- CD4 protein (marker of helper T cells) is able to interact with MHC II itsself, with invariant regions of molecule (region that isn’t polymorphic)
- CD8 protein on cytotoxic T cells can interact with MHC I proteins itsself with invariant regions of molecule
- CD4 and CD8 act as co-receptors for the TCR complex.
- Both contain Ig-like domains.
- immunoglobulin domains interact with eachother
- CD4/CD8 interact with invariant regions on MHC II/I