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Antigen Processing and
Presentation
Unit I
BT6602
1
Antigen processing and presentation
• Why is it needed?
• How does it happen?
• How are the pathways of endogenous and
exogenous antigen kept apart?
• What are the consequences?
2
T cell receptor: antigen receptor of T cells
Cannot recognise antigen in solution
Must have the antigen processed
and presented to it
Specialised molecules present
antigens to T cells:
MHC molecules
3
Antigen presentation cells required
4
T-Cell receptor
Antigen
MHC
5
MHC
6
7
Antigen Processing and Presentation
• Ag recognition by T cells REQUIRES presentation
by MHC on a cell membrane (MHC restriction)
• Pathways for Ag presentation:
a) Class I MHC associated with peptides from
endogenous Ag
b) Class II MHC associated with peptides from
exogenous Ag
8
Type of T cell activated
• MHC class I activate CD8 T cytotoxic cells
• Present endogenous (eg viral) antigens
• Target cell is killed by CD8 cell
• Any cell can become virally
infected/neoplastic
• Therefore: MHC class I is on all nucleated
cells
9
Type of T cell activated
• MHC class II activate CD4 T helper cells
• Present exogenous (eg bacteria) antigens
• CD4 T cells upregulate all immune
functions
• MHC class II found only on cells that
sample the extracellular environment
10
A question of language
• Cells that express MHC class I, present
endogenous ag
– Are killed by CD8+ cells
– Are called Target cells
• Cells that express MHC class II, present
exogenous ag
– Activate the CD4 T cell response
– Are called Antigen presenting cells
• Target cells:
– All nucleated cells express MHC class I
– Can be a target cell if virally infected/neoplastic
• Antigen presenting cells
– Limited number of cells express MHC class II
– Must sample extracellular environment
– Monocytes, MΦ, B cells
Distinctions between MHC I and MHC II
• Most cells (target cells) can
present Ag with MHC I to TC’s
• Nearly all nucleated cells
infected by MO/virus, or
abnormal proteins produced by
cancer cells, aging cells, or by
allogeneic cells from
transplants
• Associated with MHC I
requires replication of foreign
entity (i.e., abnormal protein
synthesis) within the target cell
• Only APC’s can present Ag with
MHC II to TH’s
• APC’s are of 2 categories:
– Professional APC’s
– Non-professional APC’s
• Associated with MHC II does not
require replication of entity with in
target cells
• Phagocytosis is important in Ag-
processing
MHC I MHC II
MHC
13
Ag is processed through 2 separate pathways:
*MHC I interacts with peptides from cytosolic degradation
*MHC II interacts with peptides from endocytic degradation
14
Processing Endogenous Ag:
Cytosolic pathway
• Cellular components of proteins are constantly
regulated; most have a brief half-life and are “turned
over”… the same holds true for endogenous Ag’s!
• Processing of endogenous Ag involves 3 activities:
• Peptide generation from proteolysis
• Transport to ER
• Peptide binding to MHC I
15
Endogenous Ag processing…
Peptide generation
• Proteins targeted for lysis
combine with a small protein
 ubiquitin
• Ubiquitin-protein complex
is degraded by a proteosome
• Specific proteosomes generate
peptides which can bind to
MHC I
16
Endogenous Ag processing…
Transport to ER
• Peptides from proteolysis bind to a
“transporter protein associated with
Ag processing” (TAP)
• TAP is a heterodimer which uses
ATP to help transport peptides (8-
10 aa’s) to lumen of ER
• Usually basic amino acids at COOH
end of peptide chain.
17
• MHC I assembly occurs with the aid
of chaperone proteins to promote
folding (calnexin + MHC I α chain)
• Tapasin + calreticulin brings TAP/
peptide close to MHC assembly
• Allows MHC I to bind to peptides
• MHC I-Ag exits ER to Golgi to
plasma membrane
Endogenous Ag processing…
Peptide binding to MHC I
18
Assembly and stabilization of MHC I – Ag complex
19
Processing of Exogenous Ag’s:
Endocytic pathway
• Exogenous Ag’s are typically phagocytized/
endocytized by MØ and APC’s
– Foreign Ag is degraded with in endocytic vacuole of
endocytic pathway. The pathway includes:
– Early endosomes (pH 6-6.5)
– Late endosomes or endolysosome (pH 5-6)
– Lysomes (pH 4.5 – 5)
• Ag is degraded into 13-18 aa polypeptides which bind to
MHC II
• Eventually endocytic vacuole returns to PM  recycling
surface receptors 20
Processing of Exogenous Ag’s:
Endocytic pathway
21
Processing of Exogenous Ag’s:
Manufacture of MHC II
• With in ER, α and β chains of MHC II combine with a protein – “the invariant chain” (Ii,
CD74)
• The IC binds to MHC at peptide binding cleft & then exits the ER to Golgi apparatus.
• As proteolytic activity continues, the IC is degraded to a small fragment (CLIP*)
• Another MHC II (HLA-DM (found in endosomes)) substitutes Ag for CLIP with in lysosome
• MHC II – Ag complex is transported to the PM
*CLIP = class II associated invariant chain peptide 22
Comparison of Ag-processing pathways
23
Thank you
24

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Antigen processing and presentation

  • 2. Antigen processing and presentation • Why is it needed? • How does it happen? • How are the pathways of endogenous and exogenous antigen kept apart? • What are the consequences? 2
  • 3. T cell receptor: antigen receptor of T cells Cannot recognise antigen in solution Must have the antigen processed and presented to it Specialised molecules present antigens to T cells: MHC molecules 3
  • 7. 7
  • 8. Antigen Processing and Presentation • Ag recognition by T cells REQUIRES presentation by MHC on a cell membrane (MHC restriction) • Pathways for Ag presentation: a) Class I MHC associated with peptides from endogenous Ag b) Class II MHC associated with peptides from exogenous Ag 8
  • 9. Type of T cell activated • MHC class I activate CD8 T cytotoxic cells • Present endogenous (eg viral) antigens • Target cell is killed by CD8 cell • Any cell can become virally infected/neoplastic • Therefore: MHC class I is on all nucleated cells 9
  • 10. Type of T cell activated • MHC class II activate CD4 T helper cells • Present exogenous (eg bacteria) antigens • CD4 T cells upregulate all immune functions • MHC class II found only on cells that sample the extracellular environment 10
  • 11. A question of language • Cells that express MHC class I, present endogenous ag – Are killed by CD8+ cells – Are called Target cells • Cells that express MHC class II, present exogenous ag – Activate the CD4 T cell response – Are called Antigen presenting cells
  • 12. • Target cells: – All nucleated cells express MHC class I – Can be a target cell if virally infected/neoplastic • Antigen presenting cells – Limited number of cells express MHC class II – Must sample extracellular environment – Monocytes, MΦ, B cells
  • 13. Distinctions between MHC I and MHC II • Most cells (target cells) can present Ag with MHC I to TC’s • Nearly all nucleated cells infected by MO/virus, or abnormal proteins produced by cancer cells, aging cells, or by allogeneic cells from transplants • Associated with MHC I requires replication of foreign entity (i.e., abnormal protein synthesis) within the target cell • Only APC’s can present Ag with MHC II to TH’s • APC’s are of 2 categories: – Professional APC’s – Non-professional APC’s • Associated with MHC II does not require replication of entity with in target cells • Phagocytosis is important in Ag- processing MHC I MHC II MHC 13
  • 14. Ag is processed through 2 separate pathways: *MHC I interacts with peptides from cytosolic degradation *MHC II interacts with peptides from endocytic degradation 14
  • 15. Processing Endogenous Ag: Cytosolic pathway • Cellular components of proteins are constantly regulated; most have a brief half-life and are “turned over”… the same holds true for endogenous Ag’s! • Processing of endogenous Ag involves 3 activities: • Peptide generation from proteolysis • Transport to ER • Peptide binding to MHC I 15
  • 16. Endogenous Ag processing… Peptide generation • Proteins targeted for lysis combine with a small protein  ubiquitin • Ubiquitin-protein complex is degraded by a proteosome • Specific proteosomes generate peptides which can bind to MHC I 16
  • 17. Endogenous Ag processing… Transport to ER • Peptides from proteolysis bind to a “transporter protein associated with Ag processing” (TAP) • TAP is a heterodimer which uses ATP to help transport peptides (8- 10 aa’s) to lumen of ER • Usually basic amino acids at COOH end of peptide chain. 17
  • 18. • MHC I assembly occurs with the aid of chaperone proteins to promote folding (calnexin + MHC I α chain) • Tapasin + calreticulin brings TAP/ peptide close to MHC assembly • Allows MHC I to bind to peptides • MHC I-Ag exits ER to Golgi to plasma membrane Endogenous Ag processing… Peptide binding to MHC I 18
  • 19. Assembly and stabilization of MHC I – Ag complex 19
  • 20. Processing of Exogenous Ag’s: Endocytic pathway • Exogenous Ag’s are typically phagocytized/ endocytized by MØ and APC’s – Foreign Ag is degraded with in endocytic vacuole of endocytic pathway. The pathway includes: – Early endosomes (pH 6-6.5) – Late endosomes or endolysosome (pH 5-6) – Lysomes (pH 4.5 – 5) • Ag is degraded into 13-18 aa polypeptides which bind to MHC II • Eventually endocytic vacuole returns to PM  recycling surface receptors 20
  • 21. Processing of Exogenous Ag’s: Endocytic pathway 21
  • 22. Processing of Exogenous Ag’s: Manufacture of MHC II • With in ER, α and β chains of MHC II combine with a protein – “the invariant chain” (Ii, CD74) • The IC binds to MHC at peptide binding cleft & then exits the ER to Golgi apparatus. • As proteolytic activity continues, the IC is degraded to a small fragment (CLIP*) • Another MHC II (HLA-DM (found in endosomes)) substitutes Ag for CLIP with in lysosome • MHC II – Ag complex is transported to the PM *CLIP = class II associated invariant chain peptide 22