Antigen recognition by T and B cells Role of Antigen-Presenting Cells (APC)
- Helper T cells: recognize antigen after processing and
- T and B cells exhibit fundamental differences in presentation by MHC-II on APC (dendritic cells,
macrophages, B cells).
antigen recognition
- Cytotoxic T cells: recognize antigen when it is presented on
- B cells recognize antigen free in solution (native MHC-I.
antigen). - Since most nucleated cells in the body express class I MHC,
most cells in the body can present antigen to cytotoxic T
- T cells recognize antigen after it has been cells. Although they are presenting antigen, these cells are
usually not referred to as “antigen-presenting cells”. If they
phagocytosed, degraded and small pieces of the are presenting antigen that will cause them to be killed by
antigen have been bound by MHC molecules. cytotoxic T cells, they are referred to as “target cells”.
Antigen presenting cells Professional vs Non-Professional APCs
• Remember: 1) MHC-II, 2) deliver co-stimulatory
signals
• Professional APC: DC> MΦ Φ > B cells, why?
• DC: Always express high levels of MHC-II
molecules and co-stimulatory activity (B7
molecule)
• MΦ Φ: requires activation to up-regulate MHC-II
molecules and co-stimulatory molecules (B7
molecules)
• B cells: always express MHC-II molecules but
needs to be activated to express co-stimulatory
activity (B7 molecule)
Classic
Self MHC Restriction Experiment to
show self -MHC H-2K
• Both MHC-I and MHC-II molecules can restriction
CD8 T cells
only recognize antigens when presented by
SELF-MHC molecules.
• No value for individual to have T cells
that recognize foreign antigen
associated with foreign MHC H-2K H-2K H-2b
• Self MHC restriction occurs in thymus
(-) (+) (-)
1
Role for APC?
Ag processing is required
• Classical experiment showing that B and T
cells have different requirement for antigen
recognition.
• Processing is required for Th activation
• Processing is a metabolic active process
Points Concerning Antigen Points Concerning Antigen
Processing and Presentation Processing and Presentation
1. Location of pathogen 2. Peptides derived from both self and
• viruses in cytosol, MHC class I pathway, Tc non-self proteins can associate with
response (Cytosolic pathway)
• extracellular bacteria, MHC class II MHC class I and class II molecules.
pathway, Th2 response Ab formation 3. Chemical nature of MHC groove
(Endocytic pathway)
• intracellular bacteria, MHC class II
determines which peptides it will
pathway, Th1 response cellular response bind.
(Endocytic)
MHC-I and MHC-II associated with peptides
processed in different intracellular
compartments
A) Class I MHC binds peptides derived from endogenous
antigens
B) Class I MHC binds peptides from antigens that have
been processed via the cytosolic pathway (derived
from the cytoplasm of the cell)
C) Class II MHC molecules bind peptides derived from
exogenous antigens. These antigens were
internalized by phagocytosis or endocytosis.
D) These peptides are said to have been processed
within the endocytic pathway.
2
Endogenous Pathway
• Peptides are generated by proteasome
degradation
• Peptides are transported from cytosol to the
RER
• Peptides loading onto MHC-I is aided by
chaperones
- Size
Kuby Figure 8-5 - Hydrophobicity
- The cytosolic antigen processing pathway - 2. The role of the TAP The cytosolic antigen processing pathway - 3. Assembly of the class I-peptide
(Transporter associated with Antigenic Processing). complex
1. The class I alpha chain is
- Peptides from proteasome degradation of cytoplasmic proteins are transported stabilized by calnexin.
across the membrane of the rough endoplasmic reticulum by a heterodimeric
protein designated as TAP. 2. When the alpha chain
binds beta-2-microglobulin:
- calnexin is lost
- TAP is composed of two subunits - TAP1 and TAP-2 - calreticulin and tapasin
- TAP-mediated transport is ATP-dependent bind
3. Tapasin & calreticulin
The genes for TAP1 and TAP2 are encoded within the MHC. brings the class I
molecules into the vicinity
of the TAP.
4. A cytoplasmic peptide
transported through the TAP
is loaded onto the class I
molecule.
5. Class I MHC dissociates
from calreticulin and tapasin.
Class I MHC-peptide complex
is transported to Golgi and to
Kuby Figure 8-6b
the cell surface.
Kuby Figure 8-6a
Class I MHC Pathway
Peptide is presented Class II Processing:
by MHC-I to CD8
cytotoxic T cell 1 Viral protein is made
Plasma membrane - The endocytic antigen processing pathway –
6 on cytoplasmic
ribosomes processing of externally-derived peptides
Globular viral - Antigen can be taken into cells by various means:
5
protein - intact
Peptide passes 2
phagocytosis, endocytosis, pinocytosis, receptor-
with MHC from Golgi Proteasome mediated endocytosis
body to surface rER degrades
Peptide associates protein to
with MHC-I complex peptides - Antigen taken up in these ways passes through a
series of intracellular compartments of increasing
Peptide transporter
protein moves acidity - early endosome (pH 6.5-6.0), late endosome
4 peptide into ER (pH 6.0-5.0), phagolysosome (pH <5.0)
3
MHC class I alpha
Golgi body Peptide with MHC
and beta proteins
goes to Golgi body
are made on the rER
3
The endocytic antigen processing
pathway - processing of
externally-derived peptides The endocytic antigen processing
pathway - processing of
Three major events occur in the externally-derived peptides
endosomal pathway:
The endosomal compartment is
1) Degradation of material that was completely separate from the
taken in – endosome goes through endoplasmic reticulum ---> So,
acidification and fusion with externally derived peptides are
lysosome which contain a wide usually not loaded on to MHC-I.
array of degradative enzymes
2) Loading of peptides from this
material on to class II MHC
molecules
3) Transport of class II MHC - peptide
complexes back to the cell surface
Figure 5-18 Figure 5-18
The endocytic antigen processing Invariant Chain- guides transport to
pathway - processing of endocytic Vesicles
externally-derived peptides
Class II MHC is synthesized in the ER, but
is not loaded with peptides there
because it's peptide binding site is
blocked by the invariant chain (Ii).
Once class II MHC enters the endosomal
compartments, the invariant chain is
degraded, leaving a small fragment -
CLIP - in the peptide binding site.
CLIP is removed by HLA-DM, which loads
a peptide on to class II MHC
HLA-DO inhibits HLA-DM until the cell is CLIP= Class II-associated HLA-DM – mediates exchange
activated invariant chain peptide HLA-DO – inhibits HLA-DM
Figure 5-18
Peptide MHC-II
Class II MHC Pathway Receptor (Ab)-Mediated Endocytosis
complex is presented CD4 helper T cell Globula
to CD4 helper T cell r protein
Globular protein
Endosome fuses with Endosome
Endocytosis
plasma membrane Fusion of endosome
and exocytic vesicle 1
3
Immunodominant Lysosome
peptide binds
to class II MHC 2
Exocytic vesicle fuses
with endosome Protein is processed to
Golgi releasing Ii from αβ dimer peptides in endosome
body or lysosome
Class II MHC
α Synthesis
β Ii 3 chains: α,β and Ii
Endoplasmic reticulum
4
Presentation of Non-Peptide
Antigens
• CD1 molecules (CD1a-d)
• Structurally related to MHC-I
• Encoded outside the MHC region
• Present in APC (DC>MØ>B cells)
• Presents peptides of 12-22 aa in size
• Presents to CD4, CD8 and NK cells
• Present LIPIDS and glycolipids
The End!
