Antigen processing and

presentation
Self-MHC Restriction:
T cells can only recognize Ag when it is
presented with a self-MHC molecule on
the membrane of an antigen presenting
cell (APC).
Self-MHC restriction of T
H
cells
APC T Cell
A. Rosenthal & E. Shevach
Self-MHC restriction of T
C
cells
R. Zinkernagel & P. Doherty
Role of Ag-Presenting Cells
When a primary Ab response and cell-mediated
response were induced by a protein in its native
form, a secondary Ab response could be induced
only by native Ag, whereas a secondary cell-
mediated response could be induced by either the
native or the denatured Ag.
This suggests that T cell and B cell see Ags in different ways.
Necessity of Ag Processing
(para-
Formaldehyde)
(glutaraldehyde)
Ag processing
involves the
digestion of the
protein into
peptides.
Since all cells expressing either class I
or class II MHC molecules can present
peptides to T cells, strictly speaking they
all could be designated as Ag-presenting
cells (APC).
However,..
Ag-presenting cells (APC):
Cells that display peptides associated with
class II MHC molecules to CD4
+
T
H
cells
are called APC.
Target cells:
Cells that display peptides associated with
class I MHC molecules to CD8
+
T
C
cells
are referred to as target cells.
Dendritic cells:
1. constitutively express a high level of class II MHC molecules
and costimulatory molecule
2. most effective in Ag presentation
Because nearly all nucleated cells express
class I MHC molecules, virtually any nucleated
cell is able to function as target cell presenting
endogenous Ags to CD8
+
T cells.
Most often, target cells are cells that have been
infected by a virus or some other intracellular
microorganisms.
Summary:
What is self-MHC restriction? That is, T cells can
only recognize Ag when it is presented with a self-
MHC molecule on the membrane of an antigen
presenting cell (APC).
In general, CD4
+
T
H
cells recognize antigen with class
II MHC molecules on APC and CD8
+
Tc cells
recognize antigen with class I MHC molecules on
target cells.
Antigen processing is necessary for T cell activation.
Evidence for Two Processing
and Presentation Pathways
emetine: inhibits viral protein synthesis
chloroquine: blocks the endocytic processing pathway
Endogenous Ags:
The CytosolicPathway
Peptides bound to MHC class I are
generated in the cytosol and transported
into the endoplasmic reticulum
Cytosolic proteolytic system for
degradation of intracellular proteins
Degradation of ubiquitin-protein is thought to occur in the central hollow of proteasome.
Transporters Associated with
Ag Processing (TAP)
Both TAP1 and TAP2 belong to the family
of ATP-binding cassette (ABC) proteins
found in the membranes of many cells,
including bacteria.
These proteins mediate ATP-dependent
transport of amino acids, sugars, ions, and
peptides.
Mutation of either TAP gene can prevent
antigen presentation by MHC class I
molecule.
Generation of antigenic peptides
in the cytosolic pathway
Proteosome subunits
(LMP2, LMP7, and
LMP10) favor the
production of peptides
that binds to class I
MHC molecules.

2
M
LMP and TAP genes in Mouse H-2
Class I Class I Nonclassical Nonclassical Class II Class II Class III Class III Class I Class I Nonclassical Nonclassical
LMP: 1. LMP 2 and LMP7 are encoded within MHC and induced by interferon .
2. LMP10 is not encoded within MHC and also induced by interferon .
3. LMPs preferentially generate peptides that bind to class I MHC molecules.
Such proteosomes show increased hydrolysis of peptide bonds that follow
basic and/or hydrophobic residues, the preferred anchor residues for class I.
4. Polymorphic
TAP: 1. has the highest affinity for peptides containing 8-13 amino acids.
2. favors peptides with basic and/or hydrophobic C-terminal a. a.
3. Thus, TAP is uniquely designed to transport peptides that will interact with
class I MHC molecules.
4. polymorphic
LMP and TAP genes in mouse H-2
Assembly and stabilization of
class I MHC molecules
Molecular chaperones: (promote folding of polypeptides)
calnexin: a resident membrane protein of the ER
calreticulin
tapasin (TAP-associated protein) : brings the TAP into proximity
with the class I molecules.
Summary:
There are 2 antigen processing and presentation
pathways.
Endogenous antigens are degraded into peptides in the
cytosol by proteasomes and assemble with class I MHC
molecules in the rough ER (RER).
TAP is responsible for transporting peptides from the
cytosol into the lumen of RER.
Chaperone molecules (calnexin, calreticulin, and
tapasin) help peptides assembly with class I MHC.
Exogenous Ags:
The EndocyticPathway
Peptides bound to MHC class II are
generated from internalized molecules in
endocytic vesicles.
Generation of antigenic peptides
in the endocytic pathway
40 acid-
dependent
hydrolases in
lysosomes:
1. proteases
2. nucleases
3. glycosidases
4. lipases
5. phospholipases
6. phosphatases
Assembly of class II MHC molecules
CLIP : class II-associated invariant chain peptide
a negative regulator,
inhibiting the reaction
between CLIP and
HLA-DM through binding
to HLA-DM
mediate exchange of
antigenic peptides for CLIP
Functions of invariant (Ii) chain:
1. Prevention of the binding of endogenous peptides
to class II molecules.
2. Proper folding of the class II and chains
3. Exit of class II from the RER
4. Routing of class II from the trans-Golgi to the
endocytic compartments
Class II Class II Nonclassical Nonclassical Class II Class II Class III Class III Class I Class I
Nonclassical Nonclassical
HLA-DM and HLA-DO genes
in Human HLA
Nonclassical class II genes :
DM : 1. encode class II-like molecules that facilitate the loading of
antigenic peptides into the class II MHC molecules
2. expressed within the endosomal compartments
3. not polymorphic
DO : 1. limited expression in thymus and in mature B cells
2. a regulator of class II antigen processing.
3. Not polymorphic
CLIP binds to the HLA class II molecule in a manner
identical to antigenic peptides
CLIP
Summary:
Presentation of non-peptide
antigen
T cells can respond to non-protein
antigens, such as glycolipid antigens
from Mycobacterium tuberculosis.
These non-protein antigens are
presented by members of the CD1
family of non-classical class I
molecules.
The genes encoding CD1 family of molecules
Group 1
Group 2
Crystal structure of mouse CD1 and class I molecule
1. CD1 genes can be induced by cytokines such as GM-
CSF or IL-3.
2. Antigens presented by CD1 are mycolic acid (lipid
component of the M. tuberculosis cell wall) and a
glycolipid (lipoarabinomannan) from M. leprae.
3. Expression of CD1 molecules varies according to subset;
The human CD1A, B, and C genes are expressed on
immature thymocytes and professional APC (mainly
dendritic cells). CD1C is seen on B cells.
4. The intracellular trafficking patterns of the CD1
molecules differ.
5. CD1 molecules can process antigen in TAP-deficient
cells. Together with other data, CD1 antigen presentation
seem to be through a third pathway for the processing of
antigens.
Summary:
Exogenous antigens are internalized and degraded
in the acidic endocytic compartments and
subsequently pair with class II molecules.
Peptide binding to class II molecules involves
replacing a fragment of invariant chain (CLIP) by
a process catalyzed by non-classical MHC
molecule HLA-DM.
Presentation of nonpeptide antigens (lipid and
glycolipid) derived from bacteria involves the
class I-like CD1 molecule.