ANTIGEN

PROCESSING &
PRESENTATION
Professor David Wraith

d.wraith@bham.ac.uk
Learning Objectives
■ The immune system has evolved to
combat infection
■ This has generated two arms of
cellular immunity and two paths of
antigen processing
■ Class II molecules present antigen to
CD4+ T cells and generally handle
exogenous antigens
■ This lecture will explain class II
molecule assembly, traffic and
antigen binding
■ CD8+ T cells protect against viruses
and cancer
■ The class I pathway handles intra-
cytoplasmic antigens
■ This lecture will reveal where
cytoplasmic antigens are fragmented,
how they bind class I molecules and
what controls class I MHC-peptide
association and assembly
■ Reading: Janeway’s Immunobiology
9th Edition, Chapter 6
How Are T cells
Alerted To The
Infection?
Figure 5-2
Two Primary Antigen Processing Pathways
MHC Class I MHC Class II MHC Class I MHC Class II
 MHC polymorphism
■ Proteins from individual MHC alleles can
differ from one another by up to 20 amino HLA locus Possible alleles*
acids, making each variant protein quite
distinct
HLA-A 2,884
■ Differences are localized to exposed
HLA-B 3,589
surfaces of the outer domain of the HLA-C 2,375
molecule and to the peptide-binding
groove in particular HLA-DP 16,036
■ The polymorphic residues that line the HLA-DQ 34,528
peptide-binding groove determine the HLA-DR 11,431
peptide-binding properties of
different MHC molecules *IMGT-HLA
database
Two Primary Antigen Processing Pathways

Secretory
Vesicle Golgi
Apparatus Endoplasmic
Reticulum

Endogenous
Viral & Tumour
Antigens
CLASS I & CD8
T cells
Exogenous
Antigens Nucleus
CLASS II & CD4 Endosome
T cells Lysosome
Biosynthetic Pathway of MHC Glycoproteins

■ Biosynthetic pathway of class I and II molecules diverges at or beyond the trans-

Golgi.
■ Transit of class II molecules to the cell surface is delayed substantially.
■ The biosynthetic pathway of class II molecules intersects with the endosomal route
of entry into the cell.
■ What directs class II MHC into the endocytic pathway?
Class II MHC Binds Invariant Chain

• Calnexin is a CHAPERONE
protein

• Molecular chaperones, such

as Calnexin, assist in the
folding and subunit assembly
of glycoproteins with
asparagine-linked sugar side
chains that pass through the
endoplasmic reticulum
Invariant Chain-MHC Complex
Stepwise Degradation of Ii
MHC Class II Transport and Peptide
Loading
Complex between HLA-DR and HLA-DM

A. DM stabilises empty DR. DR

residues (green) fold into P1
pocket of DR
B. Only low affinity peptide
interaction allowed due to
restricted access
C. Peptide induced conformational
change (green) in DR -> DM
dissociation
MHC class II processing and
presentation
■ Class II α- and β-chains assemble in the
endoplasmic reticulum (ER) and form a
complex with the invariant chain (Ii). The Ii–
MHC class II heterotrimer is transported
through the Golgi to the MHC class II rich
compartment of the endosomal pathway (MIIC),
either directly and/or via the plasma
membrane.
■ Endocytosed proteins and Ii are degraded by
resident proteases in the MIIC. The class II-
associated Ii peptide (CLIP) fragment of Ii
remains in the peptide-binding groove of the
MHC class II dimer and is exchanged for an
antigenic peptide with the help of the dedicated
chaperone HLA-DM (known as H2-M in mice).
MHC class II molecules are then transported to
the plasma membrane to present antigenic
peptides to CD4+ T cells.
 Two Primary Antigen Processing Pathways

Secretory
Vesicle Golgi
Apparatus Endoplasmic
Reticulum

Endogenous
Viral & Tumour
Antigens
CLASS I & CD8

Exogenous
Antigens Nucleus
CLASS II & CD4 Endosome
Lysosome
Defective Ribosomal Products

Schild &
Ramensee:
Nature (2000)
404:709-710
Proteasome from Archaebacterium
Immunoproteasome

LMP2 MECL1
b1i b2i
b1 b2
IFN-g
b7
b3

b6
b4
b5
LMP7
b5i
Constitutive Proteasome Immuno
3 proteolytic subunits b1, Proteasome
b2 & b5
Proteasome Function

■ Conventional Proteasome: Thr-dep. Protease activity cleaves after hydrophobic,

basic and acidic residues.

■ Immuno Proteasome: Formation depends on LMP7. Preference for cleavage after

hydrophobic and basic residues.
 Class I MHC Assembly Involves a Network of
ER-resident Chaperones

• Nascent Class I inserted into ER via signal recognition pore (SRP) and stabilised by interaction
with chaperone GRP78 (BiP)
• Asn86 of human class I is glycosylated. Sugar chain interacts with chaperone Calnexin
• Association with b-2m leads to calreticulin (CRT) binding
• Oxidoreductase (ERp57) and tapasin form stable unit via S-S bind and will only release class I
from TAP, for traffic to the cell surface, when a peptide of sufficient affinity has bound
But………………..

■ How are CD8 cells primed against viruses that do not infect dendritic cells?
■ There is an alternative pathway of presentation that allows exogenous antigens to
enter the class I pathway
■ This is known as ‘cross-presentation’
■ Cross-presentation is controlled by TLR signals that optimise presentation of
antigens from infectious agents
Cytotoxic T Cell Recognition
T CELL PRIMING TARGET CELL LYSIS
■ The endogenous route of ■ Most nucleated cells express class I MHC
antigen presentation is most and can present antigen by the
effective for those antigens endogenous route
expressed within dendritic
cells ■ The exogenous route of antigen
presentation is inefficient for cytotoxic T
■ The exogenous route of cell killing of target cells
antigen presentation can
result in CD8 cell activation by
DC