Dr Alok Tripathi

aquaimmuno@yahoo.co.in

+919795894495

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 What is iT?

Prior to put Antigen

before T-Cells, MHC
processed and make it
the Antigen presentable
T Cells DO
NOT recognize
antigen until &
unless it is not
put before
them by MHCs
For presentation,
antigen must be fit
into cleft of MHC
presentation zone
 Antigen Processing &
Presentation

• Preparing a peptide • The degraded

from pathogen peptide then
(antigen) which can associated with
be fix on peptide MHC, transported to
presenting cleft of membrane surface
MHC molecule and where they
displayed to TCR
Ag Antigen
Processing Presentation
 Way of Processing
Whether an antigenic
peptide associates with Ag may be processed in
class I or II molecules, is 2 ways
dictated by

the mode of entry by the site of

Exogenous Endogenous
into the cell, processing

Peptide must be
The processing is
processed inside the cell
done in endosome
(cytoplasm)

e.g. Normal cell protein,

internalize antigen by
Tumor Associated
phagocytosis,
Antigens (TAA),
endocytosis, or both
Bacteria, Virus
Experimental demonstration of self-MHC
restriction of TH cells. Peritoneal exudate
cells from strain 2, strain 13, or (2 13) F1
guinea pigs were incubated in plastic Petri
dishes, allowing enrichment of
macrophages, which are adherent cells.
The peritoneal macrophages were then
incubated with antigen. These “antigen-
pulsed” macrophages were incubated in
vitro with T cells from strain 2, strain
13, or (2 13) F1 guinea pigs, and the
degree of T-cell proliferation was assessed.
The results indicated that TH cells could
proliferate only in response to antigen
presented by macrophages that shared
MHC alleles.
 • Endogenous Antigen: Cytosolic Pathway follow the
same pathway as use in degradation of normal protein

Cytoplasmic
• Ubiquitin+ATP Proteiosome • ER
Complex • Peptide~MHC-I

• Peptides
Endogenous
Ag TAP
 A proteasome can cleave

Proteosome peptide bonds between 2 or 3

different amino acid
combinations in an ATP-
dependent process .

Degradation of ubiquitin-
protein complexes is thought
to occur within the central
hollow of the proteasome.

All three are induced by

increased levels of the T-cell
cytokine IFN-.

The peptidase activities of

proteasomes containing
LMP2, LMP7, and LMP10
preferentially generate
peptides that bind to MHC
class I molecules.
Experimental evidence indicates that the immune system
utilizes this general pathway of protein degradation to produce
small peptides for presentation with class I MHC molecules.

peptides that bind to class I MHC

molecules terminate almost exclusively
with hydrophobic or basic residues.
Subsequent experiments
showed that........
the defect in expression of
MHC in the RMA-S cell line
occurred due--------------
problem in the protein that
transports peptides from the
cytoplasm to the RER, where
class I molecules are
synthesized

When RMA-S cells were

the cells began to express class I
transfected with a functional gene
molecules on the membrane.
encoding the transporter protein,
• TAP1 and TAP2 • the TAP1 and TAP2 • these proteins
proteins each have a mediate ATP-
domain projecting into dependent
the lumen of the RER, transport of amino
and an ATP-binding acids, sugars, ions,
domain that projects and peptides.
into the cytosol.

Both TAP1 and TAP2 belong

TAP is a membrane-spanning to the family of ATP-
In addition to their multiple
heterodimer consisting of two binding cassette proteins
transmembrane segments,
proteins: found in the membranes of
many cells,

The transporter protein, designated

as TAP (transporter associated
with antigen processing)
Generation of antigenic
peptide–class I MHC
complexes
in the cytosolic
pathway.
 Process

In the cytosol,
• LMP2+ LMP7+LMP10 + a changes its catalytic
proteasome specificity to favor
Peptide + MHC –I
production of
peptides

Within the RER

membrane, then binds to
The class Iα- chain/2-β-
calreticulin and the
• a newly synthesized I α-chain microglobulin
associates with calnexin until TAP-associated protein
heterodimer
2-β-microglobulin binds to tapasin.
the chain.

When a peptide folding of MHC class I

delivered by TAP is
bound to the class Iα-
molecule,
Transported through
α-is complete and it is
the Golgi to the surface
released from the RER
of the cell.
and
 Assembly and stabilization of class I MHC molecules.

Newly formed class I chains associate with calnexin, a molecular chaperone, in the RER membrane.

Subsequent binding to 2-β microglobulin releases calnexin and allows binding to the chaperonin calreticulin and to tapasin, which is
associated with the TAP.

This association promotes binding of an antigenic peptide, which stabilizes the class I molecule–peptide complex,

allowing its release from the RER.

Exogenous Antigens: The
Endocytic Pathway
Whether an antigenic peptide associates with class I or II
molecules, is dictated by the mode of entry into the cell,
(Exo/Endo), and by the site of processing.

APC can internalize antigen by phagocytosis,

endocytosis, or both.

MΦ internalize Ag by both processes, whereas most

other APCs are not phagocytic

or are poorly phagocytic and therefore internalize

exogenous Ag only by endocytosis or pinocytosis).

e.g B cells, internalize Ag effectively by receptor-

mediated endocytosis using Ag-specific
membrane Ab as the receptor.
Peptides Are Generated from Internalized Molecules in Endocytic Vesicles

internalized Ag
takes 1–3 h to
transverse the The endocytic
endocytic pathway pathway appears
& appear at the to involve three
cell surface as increasingly acidic
Once an antigen is peptide–MHC II compartments:
internalized, it is complexes.
degraded into
peptides within
compartments of
the endocytic
processing late endosomes, or
early endosomes and lysosomes
pathway. endolysosomes
(pH 6.0–6.5); (pH 4.5–5.0).
(pH 5.0–6.0);
Generation of antigenic peptides in
the endocytic processing pathway.

Ag moves through
Internalization of
several acidic
exogenous antigen
compartments,

it is degraded into
peptides that Transported in
ultimately associate vesicles from the
with class II MHC Golgi complex.
molecules
The Invariant Chain [Ii]Guides Transport of Class II MHC
Molecules to Endocytic Vesicles

Since antigen-presenting
cells express both class I
and class II MHC
molecules, When class II MHC
molecule are
synthesized within
the RER, 3 pairs of
class II chains
associate with a
preassembled trimer
of a protein called
Some mechanism must invariant chain (Ii,
exist to prevent class II CD74).
MHC molecules from
binding to the same set of
antigenic peptides as the
class I molecules.
 invariant
chain (Ii,
CD74). preventing any
endogenously
derived peptides

+ Interact with
Peptide cleft

subsequent routing of
class II molecules to
also appears to be the endocytic
involved in the processing pathway
folding of the class from the trans-Golgi
II network.
A nonclassical class II MHC molecule called HLA-DM is required
to catalyze the exchange of CLIP with antigenic peptides.

HLA-DM is a heterodimer of and chains. However, unlike

other class II molecules, HLA-DM is not polymorphic and is
not expressed at the cell membrane but is found
predominantly within the endosomal compartment. The DM
and DM genes are located near the TAP and LMP genes in the
MHC complex of humans and DM is expressed in cells that
express classical class II molecules.
 The reaction between HLA-DM
and the class II CLIP complex
HLADO, like HLA-DM, is a
facilitating exchange of CLIP for
nonclassical and non-
another peptide is impaired in the
polymorphic class II molecule
presence of HLA-DO, which
that is also found in the MHC of
binds to HLADM and lessens the
other species.
efficiency of the exchange
reaction.

HLA-DO differs from HLA-DM in An additional difference is that

that it is expressed only by B cells the genes encoding the and the
and the thymus, and unlike other chains of HLA-DO are not
class II molecules, its expression adjacent in the MHC as are all
is not induced by IFN-. other class II and pairs