Antigen processing and presentation

Pamela Wearsch and Peter Cresswell

In antigen-presenting cells (APCs), such as dendritic cells (DCs) and B cells, heterogeneous intracellular
pathways and mechanisms are responsible for generating complexes of MHC class I and class II
molecules with peptide antigens, and complexes of CD1 molecules with lipid antigens, for presentation
to T cells. This process — referred to as antigen processing and presentation — allows T cells to
continuously assess the intracellular and extracellular milieu for signs of infection or abnormal cell
phagocytosed by APCs, this simple division is not strictly enforced. Indeed, exogenous proteins
internalized by DCs can generate peptide–MHC class I complexes that are recognized by CD8+ T cells,
a phenomenon referred to as cross-presentation. Similarly, endogenous and viral proteins can generate
peptide–MHC class II complexes that are recognized by CD4+ T cells in a process involving autophagy.
Understanding the processes and mechanisms by which antigens are captured, processed and loaded

IMMUNOLOGY growth. Although MHC class I molecules typically bind peptides derived from endogenous proteins
and MHC class II molecules typically bind peptides derived from proteins that are endocytosed or
onto MHC molecules for presentation to T cells provides us with crucial insights that are necessary for
the design of vaccines and therapeutic strategies to bolster T-cell responses.

The MHC class II pathway The MHC class I pathway

MHC class II αβ-chain dimers are assembled in the endoplasmic reticulum (ER) All nucleated cells express MHC class I molecules and present endogenous peptide
as a nonameric complex with the invariant chain (Ii), which protects against antigens to CD8+ T cells, but some DC subsets can also present exogenous
premature peptide or protein interactions in pre-lysosomal compartments. mRNA Ribosome peptides to CD8+ T cells through cross-presentation. Cell surface expression
Isolated Endogenous Viral
This complex traffics to the lysosome MHC class II compartment Exogenous of MHC class I molecules is monitored by natural killer (NK) cells, which
Autophagy membrane protein proteins
(MIIC), where Ii is subjected to sequential proteolysis. The final protein express receptors that trigger target-cell lysis when the expression of
Phagocytosis
cleavage product, a peptide known as class II-associated Ii peptide Endocytosis MHC class I is downregulated, such as occurs during some viral
Endogenous ATG8 Ub DRiP
(CLIP), occupies the peptide-binding groove and must be protein Ub Exogenous infections or cell transformation.
Ub
released prior to loading with high-affinity peptides, an protein Endogenous peptides for MHC class I presentation are generated
exchange reaction that is catalysed by the chaperone-like Autophagosome Antigen uptake Antigen-presenting cell
ER-associated in the cytosol from a variety of sources by the proteasome and
molecule HLA-DM. HLA-DO, which is expressed by B cells, increased by degradation cytosolic proteases. These epitopes and precursors are then
Early Endosome
thymic epithelial cells and certain DC subsets, can bind HLA- phagosome TLR signalling transported through the transporter associated with antigen
DM and inhibit MHC class II peptide exchange. Exogenous Retro- 26S
processing (TAP) complex into the ER, where they are further
Cross-
proteins access the MIIC through phagocytosis and translocon proteasome presentation
trimmed by ER aminopeptidases (ERAPs) to produce peptides
Misfolded
endocytosis, and endogenous proteins access the MIIC protein pathway of 8–10 amino acids. The assembly of MHC class I heavy
through autophagy. Peptide antigens of an appropriate Endosome Cytosolic
chain–β2-microglobulin (β2m) heterodimers with the short
length (~10–16 amino acids) for binding to MHC class II Late MHC class II MHC class I proteases peptides is coordinated by the peptide-loading complex,
phagosome li Phagosome
molecules are generated following reduction of disulphide ERp57 (aminopeptidases) which is composed of a disulphide-linked dimer of tapasin
bonds by interferon-γ-inducible lysosomal thiol reductase CNX and ERp57, calreticulin (CRT) and TAP molecules. Tapasin
(GILT) and cleavage by lysosomal proteases. Once ERAPs also supports peptide editing to select for the
Epitopes and
transported to the cell surface, peptide–MHC class II Peptide precursors presentation of stable peptide–MHC class I complexes
Vesicle HLA-DM trimming
complexes are presented to CD4+ T cells. on the cell surface.
Lysosome HLA-DO ATPase
Abbreviations Exogenous proteins that are internalized by some DC
Chaperone- TAP2 Retrotranslocon (p97) and macrophage subsets via phagosomes or endosomes
AEP, asparaginyl endopeptidase; AP, adaptor protein; ATG8, autophagy-
mediated
related gene 8; CNX, calnexin; DRiP, defective ribosomal product;
autophagy Nucleus TAP1 MHC class I (not shown) can be retrotranslocated (possibly involving
HSC70, heat-shock cognate protein 70; KIR, killer-cell immunoglobulin- MHC class II pathway CNX ERp57
like receptor; LAMP2, lysosomal-associated membrane protein 2; HSC70 CD1d CRT Tapasin peptide-loading components of the ER-associated degradation system)
β2m complex into the cytosol, where they are fed into the MHC class
NKR, natural-killer-cell receptor; PDI, protein disulphide isomerase; ERp57 CRT
TCR, T-cell receptor; TLR, Toll-like receptor; Ub, ubiquitin. Endolysosome I processing pathway. The minor TAP-independent
Endogenous Endosomal Lipid Peptide loading
protein ER-derived pathway is not shown for simplicity.
protease MTP and editing
S S GILT vesicle?
LAMP2 β 2m
LIP PDI
S S
Proteolytic ER
MHC cleavage MHC class I pathway
TCR class II
Cathepsins
CD4+ T cell and AEP
Vesicle MHC
CD4 class I
Cathepsin S
CLIP release No lysis
and peptide HLA-DO inhibition KIR or NK cell
Peptides CLIP
loading of HLA-DM NKR

HLA-DM release
HLA-DM (and activation?) Golgi stack

Lysosome Saposin B Trans Golgi network

or MIIC
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and gentle way to obtain highly pure cells. With no columns and no washes, EasySep® gives you The MHC class I-like CD1 molecules assemble with β2m and a lipid antigen, rather
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than a peptide antigen. Five CD1 isoforms are expressed in humans (CD1a–CD1e),
• RoboSep® (www.robosep.com), the only fully automated cell separator, performs all EasySep®
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reagents and return to separated cells in as little as 25 minutes. simplicity. Current models suggest that initial lipid binding by CD1d molecules
NKT cell occurs in the ER, possibly mediated by microsomal triglyceride transfer protein
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The RosetteSep® antibody cocktail crosslinks unwanted cells to red blood cells which are then (MTP), and then additional lipids bind through saposin-mediated exchange during Acknowledgements
removed in a standard density centrifugation step. Highly purified untouched cells are then CD1d recycling through endocytic compartments. At the cell surface, CD1d Pamela Wearsch and Peter Cresswell are Edited by Lucy Bird; copyedited by Rachel David;
available for functional assays. molecules present lipid antigens to NKT cells. at the Yale University Medical School, designed by Simon Bradbrook.
Department of Immunobiology, New Haven, © 2009 Nature Publishing Group.
For more information on the complete range of cell separation products available from STEMCELL Connecticut 06520, USA. http://www.nature.com/nri/posters/
Technologies™, please visit our website: www.stemcell.com e-mail: peter.cresswell@yale.edu antigenprocessing

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