Antigen Presentation and T Cell Stimulation by Dendritic Cells

8 Feb 2002 10:5 AR AR152-20.tex AR152-20.
SGM LaTeX2e(2001/05/10) P1: GJC

10.1146/annurev.immunol.20.100301.064828
Annu. Rev. Immunol. 2002. 20:62167

DOI: 10.1146/annurev.immunol.20.100301.064828
Copyright
c 2002 by Annual Reviews. All rights reserved
ANTIGEN PRESENTATION AND T CELL

STIMULATION BY DENDRITIC CELLS
Pierre Guermonprez1, Jenny Valladeau1, Laurence
Zitvogel2, Clotilde Thery1 and Sebastian Amigorena1
1
Institut Curie, INSERM U520, 12 rue Lhomond, 75005 Paris, France, U520,
Annu. Rev. Immunol. 2002.20:621-667. Downloaded from www.annualreviews.org
Institut Curie; e-mail: Pierre.Guermonprez@curie.fr, Jenny.Valladeau@curie.fr,

Clotilde.Thery@curie.fr, Sebastian.Amigorena@curie.fr
2
CNRS URA 1301, Laboratoire dImmunologie Cellulaire, Institut Gustave Roussy;
by University of Sussex on 06/27/12. For personal use only.
e-mail: zitvogel@igr.fr
Key Words T cell priming, immunotherapy, MHC, endocytosis, class presentation

Abstract Dendritic cells take up antigens in peripheral tissues, process them into
proteolytic peptides, and load these peptides onto major histocompatibility complex
(MHC) class I and II molecules. Dendritic cells then migrate to secondary lymphoid
organs and become competent to present antigens to T lymphocytes, thus initiating
antigen-specific immune responses, or immunological tolerance. Antigen presentation
in dendritic cells is finely regulated: antigen uptake, intracellular transport and degra-
dation, and the traffic of MHC molecules are different in dendritic cells as compared to
other antigen-presenting cells. These specializations account for dendritic cells unique
role in the initiation of immune responses and the induction of tolerance.
INTRODUCTION
Dendritic cells represent a heterogenous cell population, residing in most periph-

eral tissues, particularly at sites of interface with the environment (skin and mu-
cosae), where they represent 1%2% of the total cell numbers (1, 2). In the absence
of ongoing inflammatory and immune responses, dendritic cells constitutively pa-
trol through the blood, peripheral tissues, lymph and secondary lymphoid organs.
In peripheral tissues, dendritic cells take up self and nonself antigens. Internalized
antigens are then processed into proteolytic peptides, and these peptides are loaded
onto MHC class I and II molecules. This process of antigen uptake, degradation,
and loading is called antigen presentation.
Constitutively, however, peripheral dendritic cells present antigens quite inef-
ficiently. A signal from pathogens, often referred to as a danger signal, induces
dendritic cells to enter a developmental program, called maturation, which trans-
forms dendritic cells into efficient antigen presenting cells (APCs) and T cell acti-
vators. Bacterial and viral products, as well as inflammatory cytokines and other
self-molecules, induce dendritic cell maturation through direct interaction with
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622 GUERMONPREZ ET AL.
specific dendritic cell surface receptors. T lymphocytes, through CD40-dependent

and -independent pathways, and endothelial cells contribute to the final maturation
of dendritic cells through direct cell-to-cell contact and the secretion of cytokines
(3).
Soon after encountering a danger signal, the efficiency of antigen uptake, intra-
cellular transport and degradation, and the intracellular traffic of MHC molecules
are modified (4). Peptide loading as well as the half-life and delivery to the cell
surface of MHC molecules is increased. The surface expression of T cell costimu-
latory molecules also rises. Thus, dendritic cells become the most potent APCs, and
the only ones capable of activating naive T lymphocytes and of initiating adaptive
immune responses. To interact with T cells, however, dendritic cells also need to
migrate out of the tissues to reach secondary lymphoid organs. Concomitant with
the modifications of their antigen presentation abilities, maturation also induces
massive migration of dendritic cells out of peripheral tissues (2). Modifications
in the expression of chemokine receptors and adhesion molecules, as well as pro-

found changes of the cytoskeleton organization, contribute to the migration of
dendritic cells, through the lymph, toward secondary lymphoid organs. By linking
antigen uptake, peptide loading, and cell migration to the encounter of a danger
signal (5), dendritic cells restrict antigen presentation to those antigens internalized
during maturation, thus favoring the stimulation of T cells specific for potentially
pathogenic antigens. It is important that, in addition to presenting antigens, den-
dritic cells also influence the outcome of immune responses (2). Different dendritic
cellsubsets and dendritic cells at different maturation stages express distinct sur-
face molecules and secrete different cytokines; thus they determine selectively the
type of induced immune response.
The extraordinary efficiency of dendritic cells for T cell stimulation prompted
many groups around the world to undertake dendritic cellbased active immunother-
apy protocols (6). As the fundamental mechanisms underlying antigen presentation
in dendritic cells are being characterized, the information is used to optimize the
preparation of dendritic cells, their sensitization with antigens, and the routes of
injection. It has been fascinating in the last few years to unravel the cell biolog-
ical basis of antigen presentation in dendritic cells and to witness almost instan-
taneously how the new information and concepts have been translated into the
clinic. This review summarizes recent advances in the analysis of the cell biology
of antigen uptake and presentation, as well as T cell stimulation by dendritic cells.
We also discuss how these fundamental studies influenced the use of dendritic cells
for active immunotherapy.
ANTIGEN AND PATHOGEN RECOGNITION

AND UPTAKE IN DENDRITIC CELLS
Dendritic cells were long believed to display low endocytic and phagocytic activ-
ities. Because of their inability to take up antigens, and in spite of their high MHC
class II expression, dendritic cells were not considered as APCs. This idea lasted
ANTIGEN PRESENTATION BY DENDRITIC CELLS 623
until phagocytic Langherans cells (LCs) were shown to be precursors of some den-
dritic cells found in lymphoid organs, and bone marrow-derived dendritic cells at
an early stage of their development were shown to internalize particulate antigens
(7). Tissue dendritic cells capture pathogens, infected cells, dead cells, or their de-
rived products to use for antigen presentation. Interestingly, pathogen recognition
and uptake are in many cases accompanied by activation/maturation of dendritic
cells. Conversely, dendritic cells are also exploited by many pathogens as a route
of entry for infection.
Receptor-Mediated Endocytosis
Receptor-mediated endocytosis allows the uptake of macromolecules through spe-

cialized regions of the plasma membrane, termed coated pits. This process is
initiated by a signal in the cytoplasmic tail of the endocytic receptor, which is rec-
ognized by a family of adaptors responsible for the recruitment of clathrin lattices

and for the formation of clathrin-coated endocytic vesicles (8). A large number
of endocytic receptors are selectively expressed by subpopulations of immature
dendritic cells.
FC, COMPLEMENT, HEAT SHOCK PROTEINS, AND SCAVENGER RECEPTORS Mouse

immature dendritic cells express receptors for the Fc portion of immunoglobu-
lins (FcR): Fc RI (CD64), Fc RIII (CD16), and Fc RII (CD32) (9, 10). In hu-
man, monocyte-derived dendritic cells express mainly Fc RII (CD32) and FcR
(CD89) (11); LCs express Fc RI and FcRI (CD23) (12); and blood dendritic cells
express Fc RII and Fc RI (9). The neonatal MHC class Ilike FcR for IgG was
also found on human monocyte-derived dendritic cells (13); another receptor of
the immunoglobulin superfamily named immunoglobulin-like transcript (ILT)-3
was found in immature dendritic cells, and it allows antigen presentation (14).
Immature dendritic cells express complement receptor CR3 and CR4, but not CR1
and CR2 (15).
Heat shock proteins (hsps) derived from tumor cells or infected cells stimulate
antigen-specific T cell responses in vivo. Hsc70 and gp96 bind APCs (B cells,
macrophages, and dendritic cells) and are internalized through specific membrane
receptors (16, 17). Both binding to cells and presentation of hsp-associated peptides
were saturable and inhibited by excess of unlabelled hsp. Recently, CD91, the 2-
macroglobulin receptor, also named LRP (low density lipoproteinrelated protein),
was identified as a gp96 and hsc70 receptor on mouse APCs (18).
Scavenger receptors (SRs) are cell surface glycoproteins defined by their poten-
tial to bind chemically modified low-density lipoproteins. SRs play an important
role in host defense because they are implicated in internalization of various bac-
teria. SRs are classified according to their structure, and dendritic cells express
at least one SR: CD36 (class B-SR), involved in the uptake of apoptotic bodies
(19). Other recently identified SRs have yet to be analyzed on dendritic cells,
such as SRCL, an SR with a lectin domain, which binds Escherichia coli and
Staphylococcus aureus (20).
C-TYPE LECTIN FAMILY C-type lectins bind ligands in a Ca++-dependent manner.

They share a carbohydrate recognition domain (CRD) of 115130 amino acids,
containing four cysteins forming two disulfide bonds. Dendritic cells express sev-
eral transmembrane C-type lectins, including type I multi-CRD lectins like the
macrophage-mannose receptor (MMR) (21) or DEC205 (22) and type II single-
CRD lectins such as CD23, the low-affinity IgE receptor (12).
The MMR (mouse and human) is expressed on alveolar and differentiated
macrophages, but not on freshly isolated blood monocytes. The MMR binds sev-
eral monosaccharides as well as a wide variety of pathogen antigens, including
yeast antigens (23), lipoarabinomannan (24), and desialylated immunoglobulins
(25). The MMR is expressed on monocyte-derived dendritic cells, where it medi-
ates antigen capture, transport to endosomes and lysosomes, and efficient antigen
presentation (21). The MMR is also expressed on blood dendritic cells and inter-
stitial dendritic cells in the dermis, but not on LCs.
Mouse DEC205 was first described on mature dendritic cells, LCs, and thymic
epithelial cells (22). In contrast to the MMR, which recycles between the plasma
membrane and early endosomes, DEC205 follows an unusual intracellular path-
way after internalization, passing through late endocytic compartment (26), thanks
to a specific molecular motif (EDE) in DEC205 cytoplasmic tail, which allows ef-
ficient antigen presentation. In humans, DEC205 (also known as gp200MR6) was
only reported in blood dendritic cells, and its function is not yet documented
(27).
Dendritic cellspecific ICAM3-grabbing nonintegrin (DC-SIGN) and Langerin
are two type II lectins with mannose specificity, expressed on interstitial dendritic
cells and LCs, respectively and exclusively (28, 29). Unlike DC-SIGN, which is
more implicated in adhesion processes (see Pathogens Use Dendritic Cell Endo-
cytic Receptors), Langerin induces the formation of a unique endocytic compart-
ment of LCs, Birbeck granules (28).
Phagocytosis and Macropinocytosis

Particulate and soluble antigens are efficiently internalized by phagocytosis and
macropinocytosis, respectively. Both processes are actin dependent, require mem-
brane ruffling, and result in the formation of large intracellular vacuoles. Phagocy-
tosis is generally receptor mediated, whereas macropinocytosis is a cytoskeleton-
dependent type of fluid-phase endocytosis. In macrophages and epithelial cells,
macropinocytosis is transiently induced by growth factors or phorbol ester. In im-
mature dendritic cells, in contrast, macropinocytosis is constitutive (21). Macro-
pinocytosis represents a critical antigen uptake pathway allowing dendritic cells
to rapidly and nonspecifically sample large amounts of surrounding fluid. Phago-
cytosis, in contrast, is initiated by the engagement of specific receptors, triggering
a cascade of signal transduction, which is required for actin polymerization and
effective engulfment. In general, the same receptors mediate both phagocytosis
and clathrin-dependent endocytosis.
PHAGOCYTOSIS OF PATHOGENS Immature dendritic cells were reported to phago-

cytose almost any bacteria: cocci GRAM+ (Streptococcus aureus and Streptococ-
cus gordonii), bacilli GRAM+ (Listeria monocytogenes) and GRAM (Salmonella
typhimurium and Escherichia coli), and mycobacteria (Mycobacterium tubercu-
losis, BCG) (3). In gut epithelial monolayers, immature dendritic cells open tight
junctions, project their dendrites to the apical side of the epithelia, and sample bac-
teria therein (30). Immature dendritic cells phagocytose Saccharomyces cerevisae
and Candida albicans yeast cells and hyphae through two different pathways (coil-
ing for yeast and zipper for hyphae) (31). Finally, murine spleen dendritic cells
internalize parasites like Leishmania (L.) major, L. donovani, and L. mexicana
(32, 33).
PHAGOCYTOSIS OF APOPTOTIC AND NECROTIC BODIES Immature dendritic cells

also internalize apoptotic and necrotic bodies. In vitro, apoptosis is induced by virus
infection (influenza, vaccinia, measles, EBV, HIV1), X-ray-, - or UV-irradiation,

serum starvation, Fas- or TRAIL-ligation, or by treatment with brefeldin A. In
parallel, necrosis is generally obtained by freeze-and-thaw cycles or heating the
cells at 50 C. Human monocytederived dendritic cells internalize apoptotic and
necrotic bodies derived from T and B cell lines (34), virus-infected apoptotic
monocytes (35), or tumor cell lines, including melanoma cells (3638), squamous
cell carcinoma (39), or kidney adenocarcinoma (40). Mouse bone marrowderived
dendritic cells engulf apoptotic bodies derived from fibroblasts (41), allogenic B
cells (42), or macrophages infected with Salmonella typhimurium (43). Phagocy-
tosis of apoptotic epithelial cells by murine LCs was also observed using electron
microscopy on vaginal epithelium sections (44). Finally, immunohistology in rat
lymph node, Peyers patches, and lamina propria also revealed cytokeratin-positive
inclusions in dendritic cells, suggesting transport of apoptotic bodies by immature
dendritic cells to T cell areas of lymph nodes (45).
Molecules (solubles or receptors) mediating the engulfment of dying cells by
macrophages have been extensively analyzed. Phagocytes use complement recep-
tors, CD14, integrins (including v3 and v5, which can act in cooperation
with CD36 and thrombospondin-1), SR-family members (like CD36, lox1, or
class A-SR) (46), or the two recently described receptors PSR (47) and Mer (48).
In dendritic cells, phagocytosis of apoptotic corpses occurs through a complex
including CD36, v5 or v3, and signaling through the CrkII-Dock180-rac1
molecular complex (4951).
Regulation of Endocytosis in Dendritic Cells

Efficient antigen internalization is a specific attribute of immature dendritic cells.
During maturation, dendritic cells downregulate their endocytic capacity, thus
limiting the range of antigens that they will be able to present after leaving pe-
ripheral tissues. Downmodulation of internalization occurs through two indepen-
dent mechanisms: a decrease in cell surface expression of most antigen receptors
(e.g., MMR/FcR) and the downmodulation of both macropinocytosis and phago-

cytosis (21, 52). Recently, two independent groups have analyzed the molecular
mechanism of this process. They both found that inactivation of the small GTPases
cdc42 and rac1 blocks macropinocytosis and phagocytosis in immature dendritic
cells. In contrast to the findings of Garrett et al., West et al. could neither detect any
changes in the active forms of cdc42 during maturation nor restore macropinocy-
tosis in mature dendritic cells by microinjecting constitutively active cdc42. These
results suggest that dendritic cells may downmodulate internalization through dif-
ferent mechanisms (53, 54).
Pathogens Use Dendritic Cell Endocytic Receptors

It is interesting that pathogens learned how to use, for their own profit, unique
dendritic cell characteristicsmost notably their migratory capacity. Thus, imma-
ture dendritic cells and Langherhans cells play a critical role in the dissemination
of viruses from mucosae to draining lymph nodes. In this context, different den-
dritic cell surface molecules serve as virus, parasite, or yeast receptors. Measles
virus interacts with dendritic cells via FcRs or CD46 (55), when other viruses, like
SV40, enter cat dendritic cells through caveolae (i.e., specialized cholesterol-rich
membrane domain) (56). Parasites like Leishmania major infect LCs via CR3 (57),
and Histoplasma capsulatum yeasts are phagocytosed via very late antigen-5 (58).
The most extensively studied example of pathogen internalization in dendritic
cells is HIV. The question of the susceptibility of dendritic cells to infection by
HIV has been controversial, and dendritic cells now appear as Trojan Horses,
transporting viruses to lymph nodes for T cell infection and virus dissemination
(59). Interestingly, the HIV receptor (CD4) and coreceptor (CCR5 and CXCR4
chemokine receptors) are differentially expressed on dendritic cell subpopula-
tions (60). CCR5 (a receptor for M tropic strains) is expressed on fresh LCs,
whereas CXCR4 (a receptor for T tropic viruses) can be found on mature dendritic
cells (61). In addition, interstitial dendritic cells, but not mucosal dendritic cells,
express DC-SIGN, a surface dendritic cell molecule involved in HIV binding and
trans-infection of T cells (62). The normal function of DC-SIGN is to help den-
dritic cellT cell interactions and dendritic cell emigration out of blood or lymph
vessels through its interaction with ICAM-3 and ICAM-2, respectively (29, 63).
INDUCTION OF DENDRITIC CELL MATURATION

AND SIGNAL TRANSDUCTION
The organism bears two arms to fight pathogens: innate and adaptive immunity.
Innate immunity specifically recognizes conserved molecules of infectious anti-
gens, absent from mammalian organisms (called pathogen-associated molecular
patterns), through a family of specific receptors (pattern recognition receptors)
(64). Pathogen recognition by the immune system has two major effects. First, it
triggers effector cells of inflammation, including macrophages and polymorphonu-

clear neutrophils, which represent an immediate defense at the sites of pathogen
entry. Second, the innate immune system induces adaptive immunity. Dendritic
cells play a pivotal role as sensors of infection for the initiation of adaptive immune
responses (65).
Dendritic cells respond to two types of signals: direct recognition of pathogens
(through specific pattern-recognition receptors) and indirect sensing of infection
(through inflammatory cytokines, internal cellular compounds, and ongoing spe-
cific immune responses) (5). In response to these signals, dendritic cells are ac-
tivated to enter an integrated developmental program called maturation, which
transforms dendritic cells into efficient T cell stimulators.
In this chapter, we briefly describe the receptors involved in the induction of

dendritic cell maturation and the scarce information available concerning the signal
transduction pathways triggered by these receptors. The cell biological modifica-
tions resulting in dendritic cell migration toward secondary lymphoid organs is only
summarized. The functional consequences of dendritic cell maturation in terms of
antigen presentation and T cell stimulation is discussed in subsequent chapters.
Receptors for Dendritic Cell Activation

Five types of surface receptors were reported to trigger dendritic cell maturation:
(i) Toll-like receptors (TLR), (ii) cytokine receptors, (iii) TNF-receptor (TNF-R)
family molecules, (iv) FcR, and (v) sensors for cell death.
(i) Dendritic cells mature in response to various pathogenic compounds, inclu-
ding several bacterial wall compounds (such as lipopolysaccharideLPS),
unmethylated CpG motifs, and double-stranded RNA. Most of these mole-
cules are recognized by a large family of surface receptors called TLR
(64). In different cells of the immune system, different pathogen-associated
pattern molecules are specifically recognized by one or by a combination
of TLRs. For example, TLR4 determines responses to GRAM bacte-
ria through binding to LPS; TLR2 is involved in responses to different
GRAM+ cell wall compounds (including bacterial peptidoglycans), to bac-
terial lipoproteins, and to Klebsiella pneumoniae OmpA protein; TLR5 rec-
ognizes flagellin from both GRAM+ and GRAM bacteria; and TLR9 binds
to unmethylated CpG motifs (66). Dendritic cells express a subset of these
molecules, including TLR2, TLR3, and TLR4 (67, 68). TLR2 triggers func-
tional dendritic cell maturation in response to bacterial peptidoglycans (69),
to lipopeptides (70), to mycoplasma lipoproteins (71), and to OmpA (72).
It is interesting that TLR2 may be recruited to yeast and GRAM+ bacteria-
containing phagosomes of macrophages, which suggests that this family
of receptors may also influence endocytic and/or phagocytic functions (73).
(ii) Dendritic cells sense danger and infections indirectly through inflammatory
mediators such as TNF-, IL-1, PGE-2, whose secretion is triggered by
pathogens (2, 74).
(iii) CD4+ T cells induce dendritic cell maturation both in vivo [for the initiation
of certain cytotoxic T lymphocytes (CTLs) responses; see DC Maturation
and T Cell Priming] and in vitro. Triggering of CD40 by CD40L on T cells
induces dendritic cell maturation (75, 76), but CD40-independent mecha-
nisms also exist. Indeed, triggering of Fas and OX40L on dendritic cells by
FasL and OX40, respectively, on T cells may induce functional maturation
(77, 78) (see Do T Cells Regulate Dendritic Cell Half-Life?).
(iv) Dendritic cells may also be activated through receptors for immunoglob-
ulins (see Receptor-Mediated Endocytosis). Indeed, engagement of most
FcR, including Fc RI, Fc RIII, Fc RI, and Fc R, by immune complexes
or specific antibodies, induces dendritic cell maturation (11, 12, 79). This
process requires the FcR-associated -chain that contains an immunore-
ceptor tyrosine-containing activation motif (ITAM). Tyrosine phosphoryl-
ation of this motif may initiate dendritic cell maturation.
(v) Cell death can be sensed as a danger signal by dendritic cells. Shi et al.
showed that cell injury releases adjuvant compounds that enhance T cell
responses (80). Two studies showed that necrotic, but not apoptotic, cell
death induces mouse and human dendritic cell maturation in vitro (40, 41),
although another study found that apoptotic bodies may induce dendritic
cell maturation (81). One should be careful in interpreting these results,
however, because LPS, mycoplasma contamination (82) or CD40L ex-
pression by apoptotic bodies (83) could be responsible for dendritic cell
maturation. The nature of the dendritic cell activating compounds is still
unclear. Nucleotides, such as ATP and UTP, may activate dendritic cells
through purinergic receptors (5). Hsps including gp96, hsp90, and hsc70
are released by necrotic cells and may induce dendritic cell maturation
(84, 85). Although CD91 has recently been identified as a gp96, hsc70,
and hsp90 receptor, its role in dendritic cell activation has not yet been ex-
plored. Hsp60 and hsc70 activate macrophages through CD14 and/or TLR
(8689), but receptors engaged on dendritic cells are not characterized.
It is unlikely that all these pathways for dendritic cell maturation are redundant.
Rather, they probably result in functionally distinct dendritic cell populations. In
addition, different activation signals may act synergistically or regulate each other.
For example, human TGF--untreated dendritic cells are activated by LPS or TNF-
in contrast to TGF--treated monocyte-derived dendritic cells, which require a
CD40-dependent signal to acquire high T cell stimulating activity (90).
Signal Transduction and Dendritic Cell Maturation

Most of the receptors triggering dendritic cell maturation are expressed in other
cell types, where the signal transduction cascades have been analyzed in detail.
Few studies, however, have specifically analyzed signal transduction in dendritic
cells. In a very elegant pioneering study, Rescigno et al. showed that LPS treatment
of immature dendritic cells induces maturation through the activation of NF-B

and that it promotes dendritic cell survival by activating ERK (91). Ardeshna et al.,
more recently, confirmed that NF-B is required for LPS-induced maturation of
human monocyte-derived dendritic cells (92). They also found that survival of
these dendritic cells depends on activation of the PI3K.
CD40-mediated dendritic cell activation was also analyzed in terms of cell
signaling (93, 94). CD40 signaling in dendritic cells is initiated within plasma
membrane cholesterol-rich microdomains (rafts). TRAF2/3 and src-family pro-
tein kinases (such as Lyn) are recruited together with CD40 to membrane rafts.
TRAF2/3 initiates the activation of p38MAPK, and lyn induces ERK activation
(93). p38MAPK is required for CD40-induced IL-12 production in dendritic cells
because dendritic cells from Mkk3 (a p38 activator)-deficient mice fail to produce
IL-12 in response to CD40 stimulation (95).
Calcium-ionophore treatment can also induce phenotypical and functional mat-
uration of monocytic cells (96). Although LPS, TNF- and calcium ionophore sig-
naling pathways all lead to NF-B activation, only the calcium ionophore involves
calmodulin/calcineurin activation (96).
Recent results point at MyD88 as an essential component of TLR signaling in
dendritic cells (97). Not all TLR, however, activate a MyD88-dependent signaling
pathway. In the absence of MyD88, TLR2- and TLR9-induced dendritic cell mat-
uration is completely abolished, whereas TLR4, after LPS binding, can still induce
full dendritic cell maturation (98, 99). A MyD88-independent pathway involving
MAPK and NF-B pathways therefore exists downstream of TLR4 (98).
Finally, FcRI triggering in LCs induces phosphorylation of the tyrosine ki-
nase syk and Ca++ mobilization (12). We found recently that Fc R engage-
ment in mouse dendritic cells induces syk and ERK phosphorylation and that
syk is indispensable for immune complexinduced dendritic cell maturation
(C. Sedlik, D. Orbach, E. Schweighoffer, F. Colucci, S. Verbeek, P Ricciardi-
Castagnoli, V.L.J. Tybulewicz, J. Di Santo, S. Amigorena, in preparation). The
process of dendritic cell maturation is intimately linked to their migration out
of peripheral tissues toward lymph nodes. Dendritic cell migration is due to co-
ordinated changes in dendritic cell adhesion properties (changes in cytoskeleton
organization and integrin expression were reported) and in chemokine receptor
expression (100). Importantly, trans-endothelial migration itself participates in
dendritic cell differentiation and maturation (101). Although many studies have
analyzed dendritic cell maturation in vitro and in vivo, a coherent picture recon-
ciling all these studies with the physiological process of dendritic cell maturation
has yet to emerge. The precise coordination between the phenotypical, morpho-
logical, and functional modifications induced during maturation and the migration
of dendritic cells is unclear. It was initially proposed that only fully mature den-
dritic cells migrate efficiently. However, it is now clear that in many cases final
maturation requires dendritic cell interaction with T cells. It is unlikely that den-
dritic cells encounter specific T cells in the periphery. Thus, it is most likely that
dendritic cells leave peripheral tissues in an intermediate stage of maturation and
become fully mature in the draining lymph nodes, under the control of specific
T cells.
ANTIGEN PRESENTATION AND CROSS-PRESENTATION

CD8+ and CD4+ T cells express clonally distributed receptors that recognize
fragments of antigens (peptides) associated with MHC class I and II molecules,
respectively. Antigen degradation and peptide loading onto MHC molecules occur
intracellularly in APCs. In the last 20 years, the intracellular pathways for peptide
loading on MHC class I and II molecules were analyzed in detail. A strict compart-
mentalization of MHC class I and II biogenesis results in the loading of exogenous

and endogenous antigens on MHC class II molecules in the endocytic pathway,
and the selective loading of endogenous, but not exogenous, antigens on MHC
class I molecules in the endoplasmic reticulum (ER). This model accounts, at the
effector level, for the selective killing by MHC class Irestricted CD8+ CTLs of
virus-infected cells (expressing endogenous viral antigens), but not of neighboring
cells that have internalized inactive virus or apoptotic infected cells. This model,
however, is also in conflict with experiments from M. Bevan, who demonstrated,
over 20 years ago, that priming of CTL immune responses in vivo can occur after
presentation of exogenous antigens by MHC class I molecules (a phenomenon
that he called cross priming) (102). Recent studies in dendritic cells (and other
APCs) reconcile these two series of studies by showing that, in addition to MHC
class II, internalized antigens may also be presented by MHC class I molecules
(a phenomenon referred to as cross presentation).
MHC Class IRestricted Antigen Presentation

in Dendritic Cells
PRESENTATION OF ENDOGENOUS ANTIGENS Most peptides to be loaded on MHC
class I molecules are generated by proteasome degradation of newly synthesized
ubiquitinated proteins. The resulting peptides are transferred to the ER by special-
ized peptide transporters, TAP, and loaded on new MHC class I molecules under
the control of a loading complex composed of several ER resident chaperons (in-
cluding tapasin, calnexin, calreticulin) (103). Once associated to peptides, MHC
class I molecules are rapidly transferred through the Golgi apparatus to the plasma
membrane. Like other cells, dendritic cells present self- or virus-derived endoge-
nous antigens. Few studies have analyzed MHC class I biogenesis and endogenous
peptide loading in dendritic cells. It was shown, however, that MHC class I syn-
thesis and half-life are increased upon induction of dendritic cell maturation (104),
although less strongly than those of MHC class II (105). It is interesting that, in
contrast to MHC class II, MHC class I molecules are still efficiently synthesized
and transported to the cell surface in mature dendritic cells (104, 106), again high-
lighting the functional differences in the regulation of antigen presentation to CD4+
and CD8+ T cells. In addition, dendritic cells constitutively express low levels of
the immunoproteasome, which becomes the main type of proteasome in mature

dendritic cells (107109): LMP2, LMP7, and PA28 are all induced during den-
dritic cell maturation. This change in proteasome composition may upregulate the
efficiency of presentation of some epitopes while decreasing presentation of others
(109). Targeting of proteins for proteasomal degradation requires their ubiquitina-
tion. Interestingly, dendritic cells express a particular di-ubiquitin gene, encoded
within the MHC locus, which may participate in antigen ubiquitination (110).
WHICH INTERNALIZATION PATHWAYS LEAD TO CROSS PRESENTATION? Exogenous

antigens internalized by various pathways, however, may also be presented by
MHC class I molecules. Macropinocytosis allows receptor-independent cross pre-

sentation of soluble antigens by dendritic cells (111). Physiologically, phagocytosis
is probably a major route for antigen uptake and cross presentation. Pioneering
work in macrophages, and more recently in dendritic cells, showed that linking
antigens to latex beads, thus forcing internalization by phagocytosis, strongly in-

creased the efficiency of cross presentation (112, 113). Whether efficient cross
presentation in this case is exclusively due to phagocytosis remains unclear since
(a) a soluble antigen co-internalized with the beads was also cross presented (114)
and (b) the same antigen coupled to red blood cells, which are also phagocytosed,
did not induce cross presentation (115). These results suggest that the latex beads
could somehow disrupt endocytic compartments, thus favoring cytosol delivery
and cross presentation.
Phagocytosis of bacteria results in efficient cross presentation in both macro-
phages and dendritic cells (104, 116). Interestingly, bystander dendritic cells and
macrophages phagocytosed Salmonella-infected cells efficiently, but only den-
dritic cells cross presented bacteria-derived antigens (43). Phagocytosis of apop-
totic cells also results in efficient cross presentation of viral (34, 35, 117) and tumor
antigens (37, 38, 118). Whether apoptosis per se is required for the cross presen-
tation process is a matter of debate. Albert et al. showed that death by necrosis or
inhibitors of apoptosis resulted in inefficient cross presentation (35). Cross pre-
sentation after uptake of necrotic cells or of cellular lysates, however, may also
occur (34, 119). We found that exosomes, small membrane vesicles secreted by
tumor cells, contain tumor antigens and that their uptake by dendritic cells results
in cross presentation (120).
In addition, FcR-mediated uptake of immune complexes (79), opsonized lipo-
somes (121), or opsonized dead cells (122) promotes efficient cross presentation.
In human monocytes, antigen targeting to Fc RI also resulted in cross presentation
(123). Cross presentation after FcR-mediated uptake is observed at antigen concen-
trations 3 to 5 logs below cross presentation after fluid phase uptake (79, 121). This
cross presentation pathway is strictly dependent on immune complex receptors
since dendritic cells from receptor-deficient mice failed to cross present immune
complexes efficiently (79).
Peptides bound to different hsp, including gp96, hsp90, hsp60, and hsc70, in-
duce efficient cross presentation both in vitro and in vivo. At steady state, hsps are
loaded with endogenous peptides. Loading of gp96 with peptides, for example,
occurs in the ER and is TAP dependent (124). Hsps are released upon necrotic
cell death (84) and may be internalized by APCs through specific receptors, such
as CD91 (16, 18). Hsp-associated peptides are then somehow transferred to MHC
class I and class II molecules for antigen presentation (125, 126). Blocking hsp up-
take through CD91 prevents cross presentation, suggesting that receptor-mediated
uptake is required (125). Antigens associated to different bacterial products [in-
cluding OmpA (72), Shiga toxin (127), and the CyaA toxin (128)] too, enter cells
through specific receptors and induce efficient cross presentation.
Whether receptors only accumulate antigens within dendritic cells or whether
cross presentation requires targeting of antigens to specific endocytic compart-
ments is unknown. The wide variety of receptors that seem to promote cross
presentation could favor the first possibility. It is possible, however, that receptors
selectively drive antigens to specific endocytic cross presentation compartments.
HOW ARE INTERNALIZED ANTIGENS CROSS-PRESENTED? Two main intracellular

pathways for cross presentation were reported, resulting in either endocytic or
ER MHC class I peptide loading (129). Loading in endocytic compartments is
in general insensitive to inhibitors of protein neosynthesis (brefeldineA or cyclo-
heximide) and to inhibitors of the proteasome (lactacystine). In addition, loading
is independent of the TAP transporters and sensitive to inhibitors of lysosomal
function (ammonium chloride, chloroquine, bafilomycinA, etc.). Conversely, ER
peptide loading is blocked by proteasome inhibitors and requires the expression
of TAP. This pathway requires transport of internalized antigens from endocytic
compartments to the cytosol.
TAP-independent endocytic cross presentation requires the presence of MHC
class I molecules in the endocytic pathway. Indeed, recycling MHC class I mole-
cules are found in endosomes and lysosomes in B cells and dendritic cells (108, 130,
131). In dendritic cells, like class II molecules, MHC class I molecules are re-
distributed to the plasma membrane upon induction of maturation (108). TAP-
independent endocytic cross presentation was reported for various antigen forms
in B cells (130) and macrophages (116). In U937 cells, internalized Fc RI colo-
calizes with MHC class I molecules in rab4/5+, Lamp1 endocytic compartments,
suggesting that peptide loading may occur therein (132). Finally, endocytic load-
ing of MHC class I molecules in macrophages was directly evidenced using MHC
class I/peptide-specific antibodies (16). Therefore, it is most likely that low endo-
somal pH allows peptides generated in endosomes to exchange with endogenous
peptides bound to recycling MHC class I molecules.
TAP-dependent cross presentation was reported in dendritic cells in many
different experimental systems. Cross presentation of soluble OVA, latex bead-
bound OVA, and immunoglobulin-complexed OVA occurs through a proteasome-
sensitive, TAP-dependent pathway in mouse dendritic cells (79, 111, 133).
Cross presentation of gp96-associated peptides is also TAP dependent (18).
Hsc70-bound peptides may be presented in macrophages through TAP-dependent
or -independent pathways according to the sequence context of the peptides (16).

In this case, ER loading of the exogenous antigen was shown morphologically,
using MHC class I/peptide-specific monoclonal antibodies (16). Finally, tapasine-
deficient dendritic cells failed to cross present OVA to specific T cells, confirming
that peptide loading after antigen internalization may occur in the ER (134).
The TAP dependency of cross-presentation led K. Rock et al. to propose the
existence of a membrane transport pathway linking the lumen of endocytic com-
partments and the cytosol (133). The existence of such a transport pathway was
evidenced in macrophages (133, 135) and dendritic cells (111, 136). Indeed, den-
dritic cells bear a unique endosome-to-cytosol transport pathway that allows selec-
tive delivery of internalized antigens to the cytosol (136). Other cells like thymic
epithelial (137) or liver sinus endothelial cells (138) also present, although inef-
ficiently, soluble OVA by MHC class I in a TAP-dependent fashion. It is most
likely that, depending on the antigen, the cell type, and the route of internaliza-
tion, peptide loading may occur in endosomes, the ER, or both. It is interesting,
nevertheless, that in vivo cross priming was mostly dependent on TAP (139, 140),
suggesting that antigen delivery to the cytosol might be required for the initi-
ation of CTL immune responses in vivo (see T Cell Stimulation by Dendritic
Cells).
MHC Class IIRestricted Antigen Presentation

in Dendritic Cells
Peptide loading on MHC class II molecules occurs through a different pathway.
Soon after synthesis in the ER, three / MHC class II dimers associate to a
trimer of invariant (Ii) chains (141). These nonamers exit the ER and pass through
the Golgi apparatus before being transported to the endocytic pathway, under the
influence of transport signals present in the cytoplasmic region of the Ii chain.
Once in endosomes and lysosomes, the nonameric complexes meet an acidic,
protease-rich environment, where the Ii chain is degraded by several proteolytic
enzymes of the cathepsin family (142). MHC class II dimers become competent
to bind antigenic peptides under the control of two nonpolymorphic MHC class II
molecules HLA-DM/H2-M and HLA-DO/H2-O (in human/mouse) (143). Once
loaded with peptides, Ii chain-free MHC class II/peptide complexes reach the
plasma membrane. Antigen degradation in the endocytic pathway and the gen-
eration of antigenic peptides require several proteases, including cathepsins and
asparaginyl endopeptidases (144). Thus, MHC class II molecules bind mainly
to peptides derived from antigens present in the endocytic pathway (internalized
or membrane proteins), although many reports show endogenously synthesized
antigens presented on MHC class II molecules.
A specific feature of dendritic cells resides in the tight regulation of the cell sur-
face display of MHC class II/peptide complexes during dendritic cells life cycle.
Immature dendritic cells expose very few MHC class II/peptide complexes at their
surface. Several intracellular mechanisms cooperate to achieve this phenotype.
First, antigen degradation is inefficient in immature dendritic cells, and inter-

nalized antigens can remain intact in lysosomal compartments for several days
(145); the availability of antigenic peptides to load on MHC class II molecules
is therefore limited. Poor antigen degradation in immature dendritic cells is due
to a low efficiency of several proteases, including cathepsin B, one of the major
proteases involved in antigen degradation in dendritic cells (146). A splice variant
of Ii, Iip41, known to inhibit certain cysteine proteases (147) and detected in im-
mature dendritic cell lysosomes in association with H2-M (148), could contribute
to the poor protease activity of immature dendritic cells.
Second, even if a few antigenic peptides are formed in endosomes and lyso-
somes, very few MHC class II molecules are available to bind them in immature
dendritic cells. In some murine bone marrowderived dendritic cells, MHC class
II haplotypes with a strong affinity for Ii remain associated in lysosomes with a
partially degraded form of Ii, termed Iip10, that blocks access to the peptide bind-
ing groove (149, 150). Low protease activity of cathepsin S, the main protease for
Iip10 degradation in dendritic cells (151153), is involved here, and a role of the
cathepsin S inhibitor cystatin C, specifically localized in lysosomal compartments
together with cathepsin S, Ii, and MHC class II in immature dendritic cells, has
been proposed (150).
This type of regulation, however, is not general to all dendritic cells because it
has not been observed in immature murine dendritic cells expressing other MHC
haplotypes with lower affinity for Ii (151, 154), nor in murine dendritic cells ob-
tained from different sources or cultured in different conditions (155), nor in human
dendritic cells (105, 146). In these cases, Iip10 is degraded in lysosomes, and MHC
class II dimers then reach the cell surface in association with either antigenic pep-
tides, or the CLIP peptide derived from Ii (105, 151, 155), or without any peptide
(156). Once at the cell surface, MHC class II molecules are rapidly internalized
and can associate with new peptides in recycling endosomes before going back
to the cell surface (156, 157), or they are directed to lysosomes, where they will
finally be degraded (105, 155). This pathway results in a short-term presentation of
MHC/peptide complexes at the immature dendritic cell surface, which is probably
not sustained enough to stimulate T cells efficiently.
Maturation signals induce a coordinate modification of all these aspects of
MHC/peptide complex formation and transport. MHC class II synthesis is tran-
siently upregulated (104, 105), and protease activity of the cathepsins involved in
antigen and in Ii degradation quickly increases, maybe due to their relocalization
to more acidic compartments (146). As a result, the numbers of available peptides
and of MHC dimers free to form complexes increase concomitantly. MHC/peptide
complexes are then rapidly formed (145) and transported to endosomal vesicles
(149), where they colocalize with costimulatory (B7-2) and MHC class I molecules
before being delivered to the cell surface as clusters of molecules involved in T
cell stimulation (158). During maturation, the endocytosis activity also decreases,
and transport of internalized MHC class II molecules to lysosomes for degradation
is greatly reduced, resulting in stabilization of MHC class II/peptide complexes
at the cell surface (105, 155). Later on, MHC class II synthesis is downregulated,
and association of peptides with newly synthesized MHC molecules becomes very
inefficient, focusing the range of peptides displayed at the mature dendritic cell
surface to those arising from antigens encountered before or during the induction of
maturation. Interestingly, however, fully mature dendritic cells can still form new
MHC/peptide complexes using MHC molecules recycling from the cell surface
(157).
Anti-inflammatory cytokines can interfere with the regulation of MHC class
II processing pathways in dendritic cells. M-CSF induces a rapid and sustained
upregulation of MHC class II synthesis and transport to the cell surface in human
immature dendritic cells, but without stabilizing them at the cell surface; the re-
sulting dendritic cells are therefore very inefficient T cell stimulators (159). IL-10
inhibits the rise in protease activity induced by maturation signals by increasing the
endosomal pH, and therefore it induces a long-term inhibition of antigen process-
ing and presentation by mature dendritic cells (146). The cytokine environment of
dendritic cells, when they receive a danger signal, is therefore a crucial factor in
determining the orientation of the induced immune response.
CD1-Restricted Antigen Presentation

In addition to MHC class I and II, dendritic cells express a third class of MHC
molecules involved in antigen presentation to T cells: the CD1 proteins (160).
Five CD1 genes encoding for related nonpolymorphic proteins (CD1a, b, c, d, e)
have been identified in human, and only two in mouse, both homologues of CD1d.
On the bases of sequence homologies, CD1a, b, c, and e were classified as group
1, and CD1d as group 2 CD1 molecules. Whereas CD1a, b, and c are mainly
expressed by dendritic cells and cortical thymocytes, CD1d is expressed in various
hematopoietic cells (dendritic cells, thymocytes, circulating B cells, T cells, and
monocytes) and in intestinal epithelium. Like MHC class I, CD1 molecules are
noncovalently associated with 2-microglobulin.
Group 1 CD1 molecules, complexed with glycolipidic antigens of either en-
dogenous (self-lipids, e.g., GM1) or exogenous (e.g., mycobacteria-derived lipids)
origin, stimulate T cells of various phenotypes: CD8+ cytotoxic cells, CD4CD8
T cells, and / T cells (161). CD1/antigen association takes place mostly in
compartments of the endocytic pathway (162), where CD1 molecules are trans-
ported after internalization from the plasma membrane. Each CD1 molecule sur-
veys specific endosomal compartments: early and recycling endosomes for CD1a
(163, 164), lysosomes for CD1b, and probably all compartments for CD1c (165).
CD1a, b, and c also meet phagocytosed mycobacteria in phagosomes of different
maturation stages (166). Finally, antigen association with CD1b and CD1c can
also take place at the dendritic cell surface (167, 168).
In human and mouse, the group 2 CD1d molecule complexed with a synthetic
lipid, -galactosyl-ceramide (-GalCer), activates NKT cells, a subset of T cells
of either CD4+ or CD4CD8 phenotype that express an invariant TCR and the
NK1.1 receptor (169). The current idea is that -GalCer mimics an antigenic lipid
of cellular origin (170), which associates with CD1d in the endocytic pathway
(171). Other T cells, negative for the invariant TCR and NK1.1, recognize CD1d
molecules that have formed a complex with endogenous lipids inside the ER (171).
Very hydrophobic peptides may also form complexes with CD1d (172).
Scarce data on the specific function of CD1d expression on dendritic cells
are available. In human, NKT cells stimulated via CD1d on myeloid dendritic
cells induce dendritic cell lysis (173), thereby probably modulating the normal
immune response. In mice, CD1d-mediated interaction between NKT cells and -
GalCer-pulsed dendritic cells induces IL-12 production by dendritic cells (174) and
subsequent activation of both CD4+ and CD8+ T cells (175). We have recently
observed that stressed murine dendritic cells, because they upregulate B7, can
activate NKT cells by presenting self-antigens in a CD1d context, whereas in
resting dendritic cells, inhibitory signals delivered upon MHC class I recognition
counteract the CD1d-dependent NKT cell stimulation (176).

The fifth CD1 gene encodes for multiple isoforms of the CD1e protein, none
of which are expressed at the cell surface (177). One isoform accumulates in the
Golgi apparatus and is relocalized to late endosomes upon dendritic cell maturation
(177). The function of CD1e remains to be determined.
T CELL STIMULATION BY DENDRITIC CELLS
Priming and Tolerization

Several lines of evidence define dendritic cells as the principal professional APCs
for T cell priming. On one hand, dendritic cells were directly compared to other
cells in in vitro assays for the priming of alloreactive, nave TCR transgenic T
cells or the expansion and activation of antigen-specific nave precursors from
polyclonal populations (1). On the other hand, in vivo, injection of antigen-loaded
dendritic cells induces potent CD4+ and CD8+ T cell primary responses (178, 179).
The antigen presenting capacity of dendritic cells in situ was also assessed: Spe-
cific MHC/peptide complexes were probed (by T cells and, later on, by specific
antibodies) at the surface of the ex vivo purified dendritic cells recovered from
mice infected (180, 181) or immunized with soluble proteins in adjuvants (182
185) and DNA (186, 187). Jenkinss group directly visualized the interaction be-
tween antigen-specific transgenic T cells and antigen-loaded dendritic cells by im-
munofluorescence on lymph node sections (188). Dendritic cells were even shown
to break T cell neonatal tolerance (189), peripheral tolerance against soluble anti-
gens (190), transplantion antigens (191), peripheral tissue antigens (192, 193), and
tumor antigens (194, 195).
Importantly, dendritic cells are also involved in central and peripheral tolerance
induction. The presence of dendritic cells in the thymus medulla suggested that
they may participate in the establishment of central tolerance. Indeed, Brocker
et al. showed that the exclusive expression of MHC class II in dendritic cells
was sufficient to trigger V-specific deletion of T cells by a retrovirally encoded

superantigen (196). Using ex vivo detection of MHC/peptide complexes formed
in vivo, several studies demonstrated that thymic (197) and splenic dendritic cells
(184, 198, 199), beside B cells, are involved in antigen presentation after intra-
venous injection of tolerogenic high antigen doses. Mature dendritic cells from
the T cell areas of lymph nodes present self-antigen in the periphery (183, 200).
In addition, antibody-mediated targeting of antigen to some surface dendritic cell
molecules induced specific T cell tolerance (201, 201a). Finally, dendritic cells
could also participate in the induction of tolerance against liver allograft (202).
Cross Priming and Cross Tolerization

Shortly after the discovery of MHC restriction, M. Bevan characterized the cross
priming of CTLs against cell-associated exogenous antigens (see Antigen Presen-
tation and Cross Presentation) (102). Examples of productive immunization against

cellular antigens absent from APCs were obtained in the context of grafts, tumors,
and viruses. In the case of tumors, Huang et al. showed that (a) the APC involved in
this phenomenon derive from the bone marrow (203) and (b) the TAP transporters
are required for priming, suggesting an access of the cell-associated antigen to
the cytosol (139). TAP dependence of cross priming was recently extended to
various viral infections (140), although some epitopes bypassed presentation by
bone marrow cells or TAP requirement (204, 205). Some of the studied viruses
(like influenza) may, however, infect hematopoietic cells, bringing into question
the exclusive involvement of cross priming in the analyzed responses.
Dendritic cells are also responsible for induction of cross tolerization (i.e., the
induction of CTL tolerance against antigens that are not expressed in dendritic
cells). Kurts et al. extensively described the existence of constitutive cross presen-
tation between a model antigen expressed in the pancreas and bone marrowderived
APCs in the draining lymph nodes. This cross presentation results in transient pro-
liferation and subsequent Fas-dependent deletion of autoreactive T cells (206).
Expression of the relevant MHC class I allele selectively in dendritic cells was
sufficient to support cross tolerization (207). The implication of tumor-infiltrating
dendritic cells in cross presentation of tumor antigens has also been demonstrated
(208).
The mechanisms involved in cell-associated antigen transfer to dendritic cells
are still unclear. It is most likely that antigen uptake by dendritic cells occurs in
peripheral tissues before their migration to the draining lymph nodes. It is possi-
ble, although not demonstrated, that cross presentation requires phagocytosis of
antigen-bearing cells since cross presentation and cross priming were more effi-
cient using cell-associated antigens than using the corresponding soluble protein
(for MHC class I cross presentation) (209) or peptide (for MHC class II presenta-
tion) (42). The induction of cross presentation in low OVA-expressing mice upon
tissue destruction (by autoreactive T cells) supports this view (206). Alternative
mechanisms like capture of hsp released from necrotic cells and/or capture of
membrane vesicles (exosomes) secreted by living cells may also promote cross
presentation in vivo (see Antigen Presentation and Cross Presentation).
Lymph noderesident dendritic cells might also cross present migrating den-
dritic cells. This is suggested by the efficient presentation of antigens present on
subcutaneously injected dendritic cells by MHC class II from host-derived mature
dendritic cells in the T cell areas of the draining lymph nodes (42). Surprisingly,
however, in F1 mice immunized with parental MHC-bearing dendritic cells, the
responses are mainly restricted by the MHC allele from the injected dendritic
cells, indicating that this phenomenon does not account for efficient T cell priming
in vivo (178).
Dendritic Cell Subsets and T Cell Priming

The heterogeneity of dendritic cells in mouse and humans has been reviewed
recently (210). In mouse, as originally stated by Shortmans group, lymphoid

CD11c+CD8 +DEC205+CD11b and myeloid CD11c+CD8 DEC205
CD11b+ dendritic cells can be distinguished (211). The lymphoid versus myeloid
distinction was recently challenged by the demonstration that both common mye-
loid and lymphoid precursors can give rise to both subsets (the latter also gives rise
to LCs) (211). Splenic CD8 + and CD8 dendritic cells do not exhibit significant
differences in their ability to stimulate CD8+ nave T cells in vitro and after in vivo
intravenous injection (212). Recently, MHC/peptide complexes in splenic dendritic
cells cross presenting soluble or cell-associated OVA were analyzed (213, 214).
CD8 +CD11b dendritic cells were more efficient for MHC class I cross presen-
tation (213, 214) and CD8 CD11b+ dendritic cells for MHC class IIrestricted
presentation (213). Because the two subsets of spleen dendritic cells do not differ
strikingly in their uptake abilities, the inability of CD8 CD11b+ dendritic cells
to cross present antigen could be due to processing defects or, alternatively, to a
difference in the accessibility of the two subsets to injected antigens. This apparent
specialization of two dendritic cell subsets for MHC class I- and II-restricted anti-
gen presentation is surprising because antigen presentation through both pathways
by the same dendritic cells is required for effective CTL responses. In contradic-
tion with these results, Reis e Sousa et al. showed that intravenous administration
of lysosyme leads to MHC class II presentation mostly by CD8 +, but also by
CD8 splenic dendritic cells (185).
Subcutaneous injection of antigen-loaded CD8 + dendritic cells primed Th1
responses, whereas CD8 dendritic cells primed Th2 responses (215). These
results are consistent with the responses obtained in mice where dendritic cell
subsets were expanded by growth factor administration before immunization (216).
These studies also challenge the view that CD8 + dendritic cells induce CD4+ T
cell deletion (217). Intriguingly, subcutaneously injected CD8 + dendritic cells do
not reach draining lymph nodes efficiently, although they efficiently prime CD4+
T cells by an unknown mechanism (218).
Orientation of T cell responses by CD8 + and CD8 dendritic cells is directly
related to the cytokines produced by these dendritic cell subsets (219). The selective
induction of Th1 responses by CD8 + dendritic cells is in direct causal relationship

with their selective ability to produce IL-12 (215). Indeed, CD8 + dendritic cells
secrete more bioactive IL-12 than their CD8 counterparts; it should be noted that
IL-4 upregulates IL-12 secretion in both subsets (219). The nature of the IFN-
secreting subset remains controversial (219).
In humans, besides the GM-CSF-dependent, monocyte-derived dendritic cells
from myeloid origin, Grouard et al. characterized a CD4+CD11c dendritic cell
precursor in the T cell area of tonsils ultrastructurally as resembling plasmocytes
(220). These plasmacytoid dendritic cells (pDC or DC2), unlike monocyte-derived
dendritic cells (or DC1), are unresponsive to GM-CSF, but respond to IL-3 and
express a specific combination of surface ILT (ILT1ILT3+) (210). DC2 are also
found in thymus and blood and seem to be of lymphoid origin (210). When cul-
tured in IL-3, these dendritic cells activate Th2 cells, whereas monocyte-derived
dendritic cells activate Th1 cells (221).
Dendritic Cell Maturation and T Cell Priming

Maturation is associated with a rapid relocalization of antigen-bearing dendritic
cells to the T cell zone of secondary lymphoid organs. Maturation also enhances
surface levels of costimulatory and adhesion molecules such as B7.1 and B7.2,
which bind both CD28 and CTLA-4 on T cells. These modifications increase the
T cell priming ability of dendritic cells. If CD28 is constitutively expressed and
facilitates the initial phases of T cell priming, CTLA-4 is inducible and is involved
in the downregulation of T cell responses. Recently, it was shown that PD-1, an
inhibitory receptor expressed on T cells, binds to PD-L1/B7-H1, a member of the
B7 family expressed notably in human and murine mature dendritic cells (222).
According to Dong et al. (223), B7H-1 costimulation leads to the secretion of
IFN- and IL-10, whereas Freeman et al. (222) described a decrease in T cell
proliferation and cytokine secretion. A new dendritic cellspecific member of the
B7 family, B7-DC, was also found to bind PD-1 (224) and to efficiently costimulate
T cell proliferation, as well as IL-2 and IFN- secretion. 4-1BB, an activating T
cell costimulatory molecule, is engaged by 4-1BBL at the surface of dendritic
cells (225). 4-1BB/4-1BBL signaling leads to increased CD8+ T cell activation.
By contrast, OX40 ligation on T cells by OX40L on the dendritic cell is involved
in the differentiation of CD4+ T cells (226).
The in vivo relevance of dendritic cell maturation in T cell priming is particu-
larily clear in the case of CD8+ T cell priming. Despite several examples of direct
priming of CTL by dendritic cells in the absence of CD4+ T cells (227), several
in vivo CD8+ T cell responses are dependent on CD4+ T cell help (228). In addition,
CD4+ T cells may also convert induction of CD8+ T cell tolerance into priming,
in cases of constitutive cross presentation of peripheral self-antigens (229).
Concerning cross priming of OVA-specific CTLs after the injection of MHC-
mismatched OVA-loaded cells, Bennett et al. first demonstrated that CD4+ T cell
help required antigen recognition on the same APC (228). It was then discovered
that CD40 ligation on APCs by CD40L on CD4+ T cells was necessary and
sufficient to confer to APCs the ability to prime CTLs (230232). CD40 ligation
is sufficient to drive the maturation of dendritic cells (76) and confer on them the
ability to prime CD8+ T cells (75). The licensing effect of CD40 ligation was also
demonstrated upon injection of antigen-loaded dendritic cells (233). Moreover,
CD40 ligation can also overcome the tolerogenic effect of optimal-length peptide
administration, giving rise to a great interest in manipulating this pathway in the
field of cancer vaccines (234). This mechanism allows a temporal dissociation
of CD4+ and CD8+ antigen-specific T cell interaction with the dendritic cells:
Once licensed by CD4+ T cells, dendritic cells become competent to prime CD8+
T cells. This model bypasses the requirement for an improbable simultaneous
encounter of antigen-bearing dendritic cells with antigen-specific CD4+ and CD8+
T cells.
CD4+ T cell help may also occur in a CD4+ T celldependent, CD40-indepen-
dent manner (235), as in the case of some anti-viral responses (227), possibly by the
ligation of other receptors belonging to the TNF-R family, like RANK/TRANCE-

L (see Do T Cells Regulate DC Half-Life?). Finally, dendritic cell licensing may
occur in a CD4+ T cellindependent manner. Pathogen-derived products that drive
dendritic cell maturation, such as LPS (75) or viruses (231), can also license
dendritic cells to efficiently prime CTL responses. The expression of CD40L by
some apoptotic bodies can deliver a licensing signal that bypasses the CD4+ T cell
help for cross priming of antigens expressed by these apoptotic bodies (83).
DO IMMATURE DENDRITIC CELLS DIRECTLY TOLERIZE T CELLS? In vitro studies

show that recognition and phagocytosis by dendritic cells of apoptotic cells, which
are normally produced by the constitutive renewal of tissues, do not necessarily in-
duce dendritic cell maturation (40, 41). Induction of the chemokine receptor CCR7
upon phagocytosis of apoptotic bodies (236) suggests that, in vivo, some dendritic
cells might reach lymph nodes in a nonfully mature state following chemoat-
traction toward MIP-3 and SLC gradients. In addition, basal migratory flux of
dendritic cell from tissues to lymph nodes is observed in the absence of matura-
tion stimuli. Such constitutive dendritic cell migration was recently observed from
intestinal epithelium to the T cell area of mesenteric lymph nodes in gnotobiotic
rats (45). It was proposed that T cell recognition of self-peptide/MHC complexes
on nonfully mature dendritic cells could induce tolerance (237).
Alternatively, antigen presentation by immature dendritic cells might induce
differentiation of nave self-reactive T cells toward a suppressor/regulatory pheno-
type (238, 239). Indeed, repeated stimulations of allogenic human CD4+ T cells by
immature dendritic cells induces IL-10-producing, weakly proliferative CTLA4+
T cells (240). These cells inhibit the mature dendritic cellmediated Th1 T cell
differentiation.
In addition, injection of peptide-pulsed immature dendritic cells in healthy
volunteers specifically suppressed CD8+ memory T cell responses, through the
induction of nonlytic antigen-specific T cells secreting IL-10, low levels of IFN- ,
and no IL-4 (241). Quantification of antigen-specific T cells revealed that the loss
of effector functions was not due to a decrease in the number of circulating T

cells. A growing list of stimuli such as IL-10, glucocorticoids, vitamin D3, tumor-
derived products, or Plasmodium falciparum-infected erythrocytes, measles, or
herpes virus infection appears to inhibit dendritic cell terminal differentiation at
various stages, and in some cases, favors the induction of anergy in T cells or
the priming of regulatory Tr1 cells (secreting IL-10, but not IFN- and IL-4)
(219, 239).
In vivo, the tolerogenicity of immature dendritic cells could lead to therapeu-
tic applications because the injection of allogenic dendritic cells displaying an
immature phenotype prolongs allograft survival (242).
PLASTICITY OF THE MATURATION AND CD4+ T CELL POLARIZATION It is now clear

that the developmental origin of dendritic cells does not fully determine their ca-
pacity to polarize T cells. Other factors condition the outcome of T cell stimulation
by dendritic cells:
1. Human monocytederived dendritic cells matured by either CD40L (243),
IFN- LPS, oligo CpG nucleotides (244), or double-stranded RNA (106)
produce high levels of IL-12 and induce Th1 cells. Nevertheless, stimuli like
PGE2 may favor the differentiation of mature monocyte-derived dendritic
cells priming Th2 cells (74). The concept of polarized maturation is now
widely conforted by the growing list of factors that allow the generation of
mature IL-12-nonsecreting dendritic cells, such as PGE2, 2-agonists, TGF-
, IL-1, TNF-, and Fas engagement (74, 219). Moreover, certain dendritic
cell costimulatory molecules, such as OX40L, can deliver a Th2 polariz-
ing signal to CD4+ T cells (219). Importantly, human plasmacytoid den-
dritic cells mature and secrete high doses of IL-12 and IFN- upon viral
infection and prime Th1 differentiation (through the induction of IFN- in
human CD4+ T cells). They correspond to the previously described nat-
ural interferon producing cells (210, 245247). Finally, monocyte-derived
and plasmacytoid dendritic cells regulate each other: IFN- favors DC1 dif-
ferentiation and inhibits IL-12 production, thus promoting Th2 responses,
whereas IL-4 induces apoptosis of preDC2, thus favoring DC1differentiation
and IL-12 production (219).
2. Pathogen-derived signals appear to play a crucial role in the regulation of cy-
tokine secretion by dendritic cells. For example, Toxoplasma gondii extracts
induce IL-12 production by murine CD8 + dendritic cells in vivo and sensi-
tivity to CD40 stimulation. However, a prolonged stimulation by the extracts
paralyses IL-12 production by dendritic cells (65, 248). Candida albicans,
at the yeast stage, induces IL-12 production and Th1 protective responses
in vitro and in vivo. At the hyphae stage, the same fungus induces dendritic
cells to stimulate IL-4 secretion by T cells and no protective responses (31).
3. Tissue-specific environmental factors may participate in the phenotypic dif-
ferentiation of dendritic cells. In mouse, CD11c+ dendritic cells purified
from Peyers patches or lung, but not spleen, produce IL-4 and IL-10 and
preferentially induce in vitro Th2 polarization (249, 250).
4. Dendritic cell/T cell ratio (251) or the duration of dendritic cell stimulation
can also modulate the polarization of CD4+ T cells. Indeed, after prolonged
stimulation with LPS, IL-12 production in response to CD40L (or LPS) is
reduced. These exhausted dendritic cells prime mostly for Th2 and nonpo-
larized T cells expressing CCR7, also called central memory T cells (252).
Do T Cells Regulate Dendritic Cell Half-Life?

T cells receiving antigenic information and priming/tolerizing signals may, in turn,
regulate the half-life of antigen-bearing dendritic cells by several mechanisms. This

kind of regulation may crucially affect the dynamics of T cell priming in vivo. In-
deed, CD4+ T cell commitment to proliferation requires about 10 h of sustained
signaling in the presence of costimulation (253). In contrast, CD8+ T cells require

only a short exposure to antigen and costimulation to be committed to proliferation.
Strikingly, once committed, T cells proliferate and differentiate without the need for
further stimulation (254). The half-life of antigen-bearing dendritic cells may thus
represent a crucial parameter of T cell priming. Prolonging dendritic cell half-life
by TRANCE treatment results in increased numbers of dendritic cells reaching the
lymph nodes and increased T cell priming (255). Ex vivo antigen-loaded dendritic
cells rapidly disappear from lymph nodes after antigen-specific interaction with
nave CD4+ T cells (188). However, antigen-specific interaction between dendritic
cells and CD4+ T cells also delays LPS-induced apoptosis of splenic dendritic cells
(256). Concerning CD8+ T cells, the time course of acquisition of lytic effector
function (more rapid for memory than for nave cells) parallels the kinetics of
elimination of antigen-bearing dendritic cells (257, 258). If CTL-mediated elimi-
nation of dendritic cells limits the priming of further immune responses (257), it
does not impede the restimulation of memory CTL responses (259).
Dendritic cell half-life is regulated by T cellderived signals through:
(a) Cytokine receptors: DC2-primed Th2 cells secrete IL-4 and IL-10 that syn-
ergistically inhibits the proliferating effects of IL-3 on DC2, while IL-4
increases the maturation of monocyte-derived dendritic cells (221).
(b) MHC class II molecules: Mature dendritic cells are highly susceptible to
cell death induced by MHC class II engagement (260).
(c) RANK/TRANCE-L: Ligation by RANK-R/TRANCE promotes survival in
bone marrowderived dendritic cells, increasing their T cell stimulatory
ability (261, 262). This leads to increased survival in the lymph node of
subcutaneously injected antigen-loaded dendritic cells, thus favoring the
expansion of antigen-specific CD4+ T cells (255).
(d) TNF-R: TNF-R2 signaling plays a crucial role in the survival of LCs (263)
and bone marrowderived dendritic cells and migration of LCs (264),
whereas TNF-R1 is implicated in the NF-B-dependent maturation
process of human monocyte-derived dendritic cells (52) and murine bone

marrowderived dendritic cells (265).
(e) Death signaling receptors: Dendritic cell maturation triggers resistance to
Fas- or TRAIL-mediated death signals in correlation with increased ex-
pression of the caspase 8 inhibitory protein cFLIP (266, 267). Rescigno
et al. described a Fas-dependent pathway for the induction of maturation
in correlation with the constitutive expression of FLICE-inhibitory protein
(FLIP) (77). Another study, however, showed induction of apoptosis by
Fas ligand in immature dendritic cells (268).
( f ) The perforin/granzyme pathway: Although dendritic cells express the pro-
tease inhibitor 9 (269) and SPI-6 (270), a serpin family member, that both
inhibit granzyme B, they can be eliminated by CTLs in vitro by the perforin-
dependent pathway (258) and in vivo (257) by perforin-independent un-
known mechanisms (258). It is interesting that Th1 but not Th2 induces
SPI-6 expression in dendritic cells (270).
DENDRITIC CELLBASED IMMUNOTHERAPY
The analysis of the mechanisms underlying antigen presentation and T cell stim-
ulation provided the rationale for developing a novel strategy for immunotherapy,
based on the injection of antigen-pulsed dendritic cells. Several issues need to
be considered to ensure optimal outcome of dendritic cell-based vaccination pro-
tocols, including (a) a good manufacturing procedure used to expand dendritic
cells and (b) the source and formulation of antigen for appropriate peptide loading
on MHC molecules. These issues are still open, and only an extensive under-
standing of dendritic cell fundamental biology will allow us to develop effective
immunotherapy protocols. It is also important to consider, however, that this novel
immunotherapy approach is endowed with novel caveats and risks.
Manipulation of Dendritic Cells for Immunotherapy

EXPANDING DENDRITIC CELLS FOR THERAPEUTICAL USE Two alternative strate-
gies are currently being explored for dendritic cell expansion in immunotherapy
protocols: ex vivo and in vivo. Ex vivo, several cytokine cocktails generate im-
mature or mature monocyte-derived dendritic cells or CD34+ precursor-derived
dendritic cells from normal volunteers or patients. Some of these dendritic cell
preparations are compatible with clinical grade utilization (6). Particular attention
should be drawn toward the use of fetal calf serum (case report of anaphylaxis)
(271).
In vivo, Flt3L injected in healthy individuals elicits a profound increase in
the numbers of immature dendritic cells and precursor dendritic cells in periph-
eral blood (both DC1 and DC2). Whether this dramatic increase in dendritic cell
numbers in vivo enhances immune responses to vaccine antigens remains to be
established (272).
ANTIGEN DELIVERY TO DENDRITIC CELLS The ultimate goal is to use an anti-

genic formulation allowing a wide peptide repertoire presentation to both CD4+
and CD8+ T cells. Efficient and stable MHC class I and II presentation is ob-
tained when antigens are synthesized endogenously by the dendritic cells. This
is achieved in some malignant cells in patients with acute or chronic myeloge-
nous leukemia, which, upon cytokine stimulation, undergo differentiation into
APCs resembling dendritic cells that contain the entire repertoire of tumor anti-
gens (273275). For other diseases, the antigen must be delivered to dendritic cells
ex vivo.
Endogenous antigen expression may be obtained using bulk RNA prepared
from the tumor (276). Genetic ex vivo manipulation of dendritic cells to express
full-length antigens allows presentation of both MHC class I and II epitopes for any
MHC haplotype and the possibility to include sequences encoding immunomod-
ulators (6). The use of RNA, DNA, and substractive hybridization could allow for
enrichment and fast amplification of tumor-specific RNA (6). Universal vaccina-

tion using mRNA encoding the polypeptide component of telomerase has been
successful in eliciting tumor-specific CTLs in various tumor models, leading to
effective and broad cross protection in tumor-bearing mice (277).
Retroviral/lentiviral, adenoviral, and poxviral vectors have also been used to
elicit effective tumor antigen- and viral antigen-specific CTL responses in vitro and
in vivo (278). While retroviral vectors are suitable to transduce dividing CD34+
progenitor-derived dendritic cells (up to 70% efficiency), vaccinia or adenoviral
vectors transduce monocyte-derived dendritic cells. However, one should consider
the potential ability of the virus to interfere with the antigen presentation machinery
of the infected dendritic cells, to affect their maturation, or to cause apoptosis
(278). Regarding the problems associated with vector-specific immunity, prime-
boost strategies will have to be tested. Efficient nonviral transfection of dendritic
cells using plasmid DNA expression constructs with cationic peptide CL22 elicit
efficient antitumor responses in mice (279).
Another approach to obtain endogenous antigen synthesis has been to fuse den-
dritic cells to tumor cells. Pioneer work by Gong et al. (280), showing that murine
dendritic cells fused to colon cancer cells elicit tumor-specific CTLs in vitro and
in vivo, has been extended to several other experimental systems, confirming the
immunogenicity of such vaccines (281). Although it was postulated that dendritic
cell-tumor fusion hybrids might allow tumor antigen to gain access to the den-
dritic cell cytosol for effective MHC class I presentation, the precise mechanisms
of the immune responses leading to the clinical success of the allogeneic approach
remain to be clarified.
The alternative strategy to endogenous antigen synthesis is to provide antigens
exogenously. Peptide-pulsed bone marrow dendritic cells generate antigen-specific
T cellmediated immune responses in mice, leading to tumor eradication (179,
282284). Pulsing synthetic peptides derived from known tumor (285287) or
viral (288) antigen precursors onto human ex vivogenerated dendritic cells elicited
peptide-specific CTL responses in vitro. These peptides, however, only reside on
the dendritic cell surface for short periods of time (289, 290), are limited in use
for patients bearing the appropriate MHC haplotypes, and were associated with
antigen and/or MHC class I loss variants in vivo (291, 292).
As mentioned in MHC Class I Restricted Antigen Presentation in Dendritic
Cells, apoptotic cells also allow efficient antigen loading in dendritic cells. Nouri-
Shirazi et al. (118) and Berard et al. (38) have shown that dendritic cells loaded with
killed allogeneic melanoma or prostate cells prime nave T cells to differentiate
into CTLs specific for a broad spectrum of shared tumor antigens and recognizing
naturally processed antigens.
We have shown that a novel tumor cell secretory compartment, exosomes, is a
defined source of shared rejection tumor antigens released in membrane vesicles
by living tumor cells. Tumor-derived exosomes contain whole native cytosolic
and/or endosomal tumor antigens and constitutive hsps. Exosomes transfer tumor
antigen to dendritic cells and induce peptide-specific, MHC class Irestricted cross
presentation to T cell clones and in vitro tumor-specific CTL responses in patients
lymphocytes (renal cell cancer and melanoma). Moreover, exosomes promote cross
protecting antitumor effects in both syngeneic and allogeneic mouse tumor models,
suggesting that they may transfer shared tumor antigens (120).
Potential Caveats of Dendritic CellBased

Immunotherapy of Cancer
Activated dendritic cells can break tumor-induced tolerance. Tumor antigens in-
clude tissue-specific antigens such as melanoma/melanocyte antigens and antigens
widely expressed in peripheral normal tissues, albeit at low levels. To use such tu-
mor antigens for immunotherapy, one should consider the risk of autoimmunity. For
example, dendritic cells pulsed with a peptide from a natural melanocyte/melanoma
tumor antigen could induce T celldependent autoimmune responses, leading to
melanoma rejection and vitiligo due to autoimmunity at the vaccination site (194).
In mice expressing LCMV antigen in the pancreas, efficient dendritic cell vacci-
nation against a LCMV-expressing tumor led to overt diabetes (195).
In humans, a link between dendritic cellmediated cross presentation and au-
toimmunity was suggested by the analysis of patients with autoimmune paraneo-
plastic cerebellar degeneration (PCD), who have limited breast or ovarian cancer
evolution, and an expanded population of anti-cdr2 CTLs (cdr2 is normally ex-
pressed in neurons and testis) (293). Dendritic cells from PCD patients phagocy-
tosed apoptotic tumor lines expressing cdr2 and induced potent anti-cdr2 cytolysis
from autologous T cells. In PCD, cross presentation of apoptotic tumor cells by
dendritic cells might provide the initial stimulus for CTLs in vivo.
Another potential problem is that, if cross presentation allows the priming of
protective CTL responses against tumors or infectious agents, it could also result
in the priming of CTLs specific for antigenic structures that are not presented by
infected or tumoral target cells and, thus, are devoid of protective ability (294, 295).
Conversely, dendritic cell processing of tumoral antigens by the immunoprotea-
some can impede the presentation of some epitopes otherwise presented in tumoral
cells devoid of immunoproteasome (109).
Human Trials
The first controlled evidence of the adjuvant role of mature dendritic cells in normal
volunteers was brought up in 1999/2000 by Steinmans group (296). While injec-
tion of unpulsed dendritic cells or antigen alone (KLH, TT, MP) failed to immunize,
priming of CD4+ T cells to KLH, boosting of TT specific T cell immunity, and
enhanced precursor frequency of MP-specific IFN- producing CTLs were shown
following injection of a single dose of mature dendritic cells. When T cells were
boosted in culture, there was an increase in MHC-specific tetramer-binding cells
and CTL after dendritic cell vaccination. Responses to all three antigens peaked at
3090 days after immunization and declined thereafter. Three volunteers were rein-
jected with mature dendritic cells pulsed with matrix protein alone and displayed
a rapid boost in matrix protein-specific immunity (faster and greater magnitude of
CD8+ T cell specific responses), with direct cytolytic activity in one case.
Clinical trials using dendritic cell-based immunotherapy have been initiated or

completed in melanoma, lymphoma, myeloma, prostate, and renal cancer patients
(297). All published trials demonstrated safety. In most of these studies, imma-
ture monocyte-derived dendritic cells pulsed with tumor antigen peptides were
used (239, 298300). The recent observation (see Dendritic Cell Maturation and
T Cell Priming) that injection of immature dendritic cells in human suppresses
CTL responses (241) suggests, however, that mature dendritic cells could be a
better antitumor adjuvant. A series of clinical trials based on the blood cell sort-
ing of dendritic cells upfront (301) or following Flt3L therapy (so-called mature
dendritic cells after two days in culture for antigen loading) was recently pub-
lished (272, 302, 303). While Flt3L directly injected in metastatic patients was
mediating dendritic cell expansion, no objective clinical response was seen (some
enhanced DTH responses to common antigens and tumor dendritic cells infiltrates
were described). Using monocyte-derived, peptide-pulsed dendritic cells fully ma-
tured under rigorous quality control parameters (GM-CSF/IL4 followed by IL1,
TNF, PGE2, IL-6), two groups (304, 305) reported clinical and immunological
responses in melanoma or renal cancer patients, specially in those with low tumor
burden. CD34+-derived dendritic cells are also being used in clinical trials (306).
A provocative report from Kugler et al. (307) recently described therapy of
17 renal cell carcinoma patients with tumor celldendritic cell hybrids (electrofu-
sion). Dendritic cells were allogeneic, whereas tumor cells were autologous. The
mean follow up was 13 months, and 4 complete responses, 2 partial responses,
and 1 minor response were achieved on metastatic diseases. They also demon-
strated recruitment of CD8+ T lymphocytes into tumor sites. Fusions and hybrid
formation were documented only in 10%15% of total reinjected cells.
Safety and antigen presenting properties of allogeneic (HLA-A2, pulsed with
Gal, Pol, Env peptides) or autologous dendritic cells were addressed in 6 HLA-
A2+, HIV infected patients. Both dendritic cell infusions were well tolerated, and
in patients with normal CD4+ T cell counts, administration of these antigen-pulsed
cells enhanced the immune response to HIV with no effect on the viral load (308).
They further showed persistance of donor cells following adoptive transfer of

allogeneic dendritic cells.
Besides antigen-pulsed dendritic cells injection in patients, alternative strategies
could rely on direct expansion of dendritic cells in patients or on targeting antigens
to dendritic cells in vivo. Hsps could represent such a vehicle to deliver peptides
to dendritic cells. Alternatively, we showed that, like tumor cells, murine dendritic
cells secrete antigen presenting vesicles of endosomal origin, called exosomes
(309). Dendritic cellderived exosomes induce potent antitumor immune responses
and eradication. They have a unique protein composition, distinct from other cel-
lular compartments, including apoptotic microvesicles (310, 311). Peptide-loaded
exosomes are efficiently presented by murine dendritic cells to CD4+ T cells in
vitro and can stimulate nave T cells in vivo (C. Thery, L. Duban, P. Veron, O. Lantz,
S. Amigorena, et al., in preparation). Immature human dendritic cells also secrete
exosomes of similar protein composition and function (309). A good manufac-
turing process has been engineered allowing scale up of exosome production and
purification with safety profile of the product in mice. A clinical trial has recently
been launched, aimed at vaccinating subcutaneously metastatic melanoma and
inoperable lung cancer patients with autologous dendritic cellderived exosomes
pulsed with Mage3 class I and II peptides.
Ideal vaccination processes should be cell free, off-shelf reagents, allow man-
ufacturing in large scale, and use immunorelevant tumor antigens regardless of
patient HLA haplotype. We are, obviously, far from this goal. In spite of encour-
aging preliminary clinical results in pilot studies and phase I/II trials (demonstrat-
ing feasibility and safety), dendritic cellbased immunotherapy awaits firm proof
of efficacy. In particular, correlations between enhanced specific CTL precursor
frequency and clinical responses have not been reported yet.
ACKNOWLEDGMENTS
P.G. was supported by Association pour la Recherche contre le Cancer, J.V. by
Ligue Nationale contre le Cancer, C.T. by Fondation pour la Recherche Medicale.
Due to space limitation, we could only cite a fraction of the published work, which
does not undermine the great value of uncited studies.
Visit the Annual Reviews home page at www.annualreviews.org
LITERATURE CITED
1. Banchereau J, Steinman RM. 1998. Den- dritic cells. Annu. Rev. Immunol. 18:767
dritic cells and the control of immunity. 811
Nature 392:24552 3. Bell D, Young JW, Banchereau J. 1999.
2. Banchereau J, Briere F, Caux C, Davoust Dendritic cells. Adv. Immunol. 72:255
J, Lebecque S, Liu YJ, Pulendran B, 324
Palucka K. 2000. Immunobiology of den- 4. Thery C, Amigorena S. 2001. The cell
biology of antigen presentation in den- Fc receptor for IgG is functionally ex-

dritic cells. Curr. Opin. Immunol. 13:45 pressed in monocytes, intestinal macro-
51 phages, and dendritic cells. J. Immunol.
5. Gallucci S, Matzinger P. 2001. Danger 166:326676
signals: SOS to the immune system. Curr. 14. Cella M, Dohring C, Samaridis J, Dess-
Opin. Immunol. 13:11419 ing M, Brockhaus M, Lanzavecchia A,
6. Nestle FO, Banchereau J, Hart D. 2001. Colonna M. 1997. A novel inhibitory re-
Dendritic cells: on the move from bench ceptor (ILT3) expressed on monocytes,
to bedside. Nat. Med. 7:76165 macrophages, and dendritic cells involved
7. Steinman RM, Swanson J. 1995. The en- in antigen processing. J. Exp. Med. 185:
docytic activity of dendritic cells. J. Exp. 174351
Med. 182:28388 15. Reis e Sousa C, Stahl PD, Austyn JM.
8. Slepnev VI, De Camilli P. 2000. Acces- 1993. Phagocytosis of antigens by Lan-

sory factors in clathrin-dependent synap- gerhans cells in vitro. J. Exp. Med. 178:
tic vesicle endocytosis. Nat. Neurosci. 50919
Rev. 1:16172 16. Castellino F, Boucher PE, Eichelberg K,

9. Fanger NA, Wardwell K, Shen L, Ted- Mayhew M, Rothman JE, Houghton AN,
der TF, Guyre PM. 1996. Type I (CD64) Germain RN. 2000. Receptor-mediated
and type II (CD32) Fc gamma receptor- uptake of antigen/heat shock protein com-
mediated phagocytosis by human blood plexes results in major histocompatibility
dendritic cells. J. Immunol. 157:54148 complex class I antigen presentation via
10. Esposito-Farese ME, Sautes C, de la two distinct processing pathways. J. Exp.
Salle H, Latour S, Bieber T, de la Salle Med. 191:195764
C, Ohlmann P, Fridman WH, Cazenave 17. Arnold-Schild D, Hanau D, Spehner D,
JP, Teillaud JL, et al. 1995. Membrane and Schmid C, Rammensee HG, de la Salle H,
soluble Fc gamma RII/III modulate the Schild H. 1999. Cutting edge: receptor-
antigen-presenting capacity of murine mediated endocytosis of heat shock pro-
dendritic epidermal Langerhans cells teins by professional antigen-presenting
for IgG-complexed antigens. J. Immu- cells. J. Immunol. 162:375760
nol. 155:172536 18. Basu S, Binder RJ, Ramalingam T, Sri-
11. Geissmann F, Launay P, Pasquier B, vastava PK. 2001. CD91 is a common
Lepelletier Y, Leborgne M, Lehuen A, receptor for heat shock proteins gp96,
Brousse N, Monteiro RC. 2001. A sub- hsp90, hsp70, and calreticulin. Immunity
set of human dendritic cells expresses IgA 14:30313
Fc receptor (CD89), which mediates in- 19. Platt N, da Silva RP, Gordon S. 1998.
ternalization and activation upon cross- Recognizing death: the phagocytosis of
linking by IgA complexes. J. Immunol. apoptotic cells. Trends Cell Biol. 8:365
166:34652 72
12. Jurgens M, Wollenberg A, Hanau D, de 20. Nakamura K, Funakoshi H, Miyamoto K,
la Salle H, Bieber T. 1995. Activation of Tokunaga F, Nakamura T. 2001. Molec-
human epidermal Langerhans cells by en- ular cloning and functional characteriza-
gagement of the high affinity receptor for tion of a human scavenger receptor with
IgE, Fc epsilon RI. J. Immunol. 155:5184 C-type lectin (SRCL), a novel member
89 of a scavenger receptor family. Biochem.
13. Zhu X, Meng G, Dickinson BL, Li X, Biophys. Res. Commun. 280:102835
Mizoguchi E, Miao L, Wang Y, Robert C, 21. Sallusto F, Cella M, Danieli C, Lanzavec-
Wu B, Smith PD, Lencer WI, Blumberg chia A. 1995. Dendritic cells use macro-
RS. 2001. MHC class I-related neonatal pinocytosis and the mannose receptor
to concentrate macromolecules in the 29. Geijtenbeek TB, Krooshoop DJ, Bleijs

major histocompatibility complex class DA, van Vliet SJ, van Duijnhoven GC,
II compartment: downregulation by cy- Grabovsky V, Alon R, Figdor CG, van
tokines and bacterial products. J. Exp. Kooyk Y. 2000. DC-SIGN-ICAM-2 inter-
Med. 182:389400 action mediates dendritic cell trafficking.
22. Jiang W, Swiggard WJ, Heufler C, Peng Nat. Immunol. 1:35357
M, Mirza A, Steinman RM, Nussenz- 30. Rescigno M, Urbano M, Valzasina B,
weig MC. 1995. The receptor DEC-205 Francolini M, Rotta G, Bonasio R, Gra-
expressed by dendritic cells and thymic nucci F, Kraehenbuhl JP, Ricciardi-
epithelial cells is involved in antigen pro- Castagnoli P. 2001. Dendritic cells ex-
cessing. Nature 375:15155 press tight junction proteins and penetrate
23. Ezekowitz RA, Williams DJ, Koziel H, gut epithelial monolayers to sample bac-
Armstrong MY, Warner A, Richards FF, teria. Nat. Immunol. 2:36167

Rose RM. 1991. Uptake of Pneumocystis 31. dOstiani CF, Del Sero G, Bacci A, Mon-
carinii mediated by the macrophage man- tagnoli C, Spreca A, Mencacci A, Ric-
nose receptor. Nature 351:15558 ciardi-Castagnoli P, Romani L. 2000.

24. Prigozy TI, Sieling PA, Clemens D, Ste- Dendritic cells discriminate between
wart PL, Behar SM, Porcelli SA, Bren- yeasts and hyphae of the fungus Can-
ner MB, Modlin RL, Kronenberg M. dida albicans. Implications for initia-
1997. The mannose receptor delivers li- tion of T helper cell immunity in vitro
poglycan antigens to endosomes for pre- and in vivo. J. Exp. Med. 191:1661
sentation to T cells by CD1b molecules. 74
Immunity 6:18797 32. Gorak PM, Engwerda CR, Kaye PM.
25. Dong X, Storkus WJ, Salter RD. 1999. 1998. Dendritic cells, but not macro-
Binding and uptake of agalactosyl IgG phages, produce IL-12 immediately fol-
by mannose receptor on macrophages and lowing Leishmania donovani infection.
dendritic cells. J. Immunol. 163:542734 Eur. J. Immunol. 28:68795
26. Mahnke K, Guo M, Lee S, Sepulveda H, 33. Konecny P, Stagg AJ, Jebbari H, Eng-
Swain SL, Nussenzweig M, Steinman lish N, Davidson RN, Knight SC. 1999.
RM. 2000. The dendritic cell receptor Murine dendritic cells internalize Leish-
for endocytosis, DEC-205, can recycle mania major promastigotes, produce IL-
and enhance antigen presentation via ma- 12 p40 and stimulate primary T cell prolif-
jor histocompatibility complex class II- eration in vitro. Eur. J. Immunol. 29:1803
positive lysosomal compartments. J. Cell 11
Biol. 151:67384 34. Subklewe M, Paludan C, Tsang M, Mah-
27. Kato M, Neil TK, Fearnley DB, McLel- nke K, Steinman R, Munz C. 2001.
lan AD, Vuckovic S, Hart DN. 2000. Ex- Dendritic cells cross-present latency
pression of multilectin receptors and com- gene products from Epstein-Barr virus-
parative FITC-dextran uptake by human transformed B cells and expand tumor-
dendritic cells. Int. Immunol. 12:151119 reactive CD8(+) killer T cells. J. Exp.
28. Valladeau J, Ravel O, Dezutter-Dambu- Med. 193:40512
yant C, Moore K, Kleijmeer M, Liu Y, 35. Albert ML, Sauter B, Bhardwaj N. 1998.
Duvert-Frances V, Vincent C, Schmitt D, Dendritic cells acquire antigen from apop-
Davoust J, Caux C, Lebecque S, Saeland totic cells and induce class I-restricted
S. 2000. Langerin, a novel C-type lectin CTLs. Nature 392:8689
specific to Langerhans cells, is an endo- 36. Shaif-Muthana M, McIntyre C, Sisley K,
cytic receptor that induces the formation Rennie I, Murray A. 2000. Dead or alive:
of Birbeck granules. Immunity 12:7181 immunogenicity of human melanoma
cells when presented by dendritic cells. bystander dendritic cells. J. Exp. Med.
Cancer Res. 60:644147 191:61324
37. Russo V, Tanzarella S, Dalerba P, Rigatti 44. Parr MB, Kepple L, Parr EL. 1991.
D, Rovere P, Villa A, Bordignon C, Tra- Langerhans cells phagocytose vaginal ep-
versari C. 2000. Dendritic cells acquire ithelial cells undergoing apoptosis during
the MAGE-3 human tumor antigen from the murine estrous cycle. Biol. Reprod.
apoptotic cells and induce a class I- 45:25260
restricted T cell response. Proc. Natl. 45. Huang FP, Platt N, Wykes M, Major JR,
Acad. Sci. USA 97:218590 Powell TJ, Jenkins CD, MacPherson GG.
38. Berard F, Blanco P, Davoust J, Neidhart- 2000. A discrete subpopulation of den-
Berard EM, Nouri-Shirazi M, Taquet N, dritic cells transports apoptotic intestinal
Rimoldi D, Cerottini JC, Banchereau J, epithelial cells to T cell areas of mesen-
Palucka AK. 2000. Cross-priming of teric lymph nodes. J. Exp. Med. 191:435
naive CD8 T cells against melanoma anti- 44
gens using dendritic cells loaded with 46. Savill J, Fadok V. 2000. Corpse clearance
killed allogeneic melanoma cells. J. Exp. defines the meaning of cell death. Nature
Med. 192:153544 407:78488
39. Hoffmann TK, Meidenbauer N, Dwo- 47. Fadok VA, Bratton DL, Rose DM, Pear-
racki G, Kanaya H, Whiteside TL. 2000. son A, Ezekewitz RA, Henson PM. 2000.
Generation of tumor-specific T-lympho- A receptor for phosphatidylserine-spe-
cytes by cross-priming with human den- cific clearance of apoptotic cells. Nature
dritic cells ingesting apoptotic tumor 405:8590
cells. Cancer Res. 60:354249 48. Scott RS, McMahon EJ, Pop SM, Reap
40. Sauter B, Albert ML, Francisco L, EA, Caricchio R, Cohen PL, Earp HS,
Larsson M, Somersan S, Bhardwaj N. Matsushima GK. 2001. Phagocytosis and
2000. Consequences of cell death: ex- clearance of apoptotic cells is mediated by
posure to necrotic tumor cells, but not MER. Nature 411:20711
primary tissue cells or apoptotic cells, in- 49. Albert ML, Kim JI, Birge RB. 2000.
duces the maturation of immunostimula- alphavbeta5 integrin recruits the CrkII-
tory dendritic cells. J. Exp. Med. 191:423 Dock180-rac1 complex for phagocytosis
34 of apoptotic cells. Nat. Cell Biol. 2:899
41. Gallucci S, Lolkema M, Matzinger P. 905
1999. Natural adjuvants: endogenous ac- 50. Albert ML, Pearce SF, Francisco LM,
tivators of dendritic cells. Nat. Med. 5: Sauter B, Roy P, Silverstein RL, Bhard-
124955 waj N. 1998. Immature dendritic cells
42. Inaba K, Turley S, Yamaide F, Iyoda T, phagocytose apoptotic cells via alphav-
Mahnke K, Inaba M, Pack M, Sub- beta5 and CD36, and cross-present anti-
klewe M, Sauter B, Sheff D, Albert M, gens to cytotoxic T lymphocytes. J. Exp.
Bhardwaj N, Mellman I, Steinman RM. Med. 188:135968
1998. Efficient presentation of phagocy- 51. Rubartelli A, Poggi A, Zocchi MR.
tosed cellular fragments on the major his- 1997. The selective engulfment of apop-
tocompatibility complex class II products totic bodies by dendritic cells is medi-
of dendritic cells. J. Exp. Med. 188:2163 ated by the alpha(v)beta3 integrin and
73 requires intracellular and extracellular
43. Yrlid BU, Wick MJ. 2000. Salmonella- calcium. Eur. J. Immunol. 27:1893900
induced apoptosis of infected macro- 52. Sallusto F, Lanzavecchia A. 1994. Effi-
phages results in presentation of a cient presentation of soluble antigen
bacteria-encoded antigen after uptake by by cultured human dendritic cells is
maintained by granulocyte/macrophage 61. Zaitseva M, Blauvelt A, Lee S, Lapham

colony-stimulating factor plus interleukin CK, Klaus-Kovtun V, Mostowski H,
4 and downregulated by tumor necrosis Manischewitz J, Golding H. 1997. Ex-
factor alpha. J. Exp. Med. 179:110918 pression and function of CCR5 and
53. West MA, Prescott AR, Eskelinen EL, CXCR4 on human Langerhans cells and
Ridley AJ, Watts C. 2000. Rac is required macrophages: implications for HIV pri-
for constitutive macropinocytosis by den- mary infection. Nat. Med. 3:136975
dritic cells but does not control its down- 62. Geijtenbeek TB, Kwon DS, Torensma
regulation. Curr. Biol. 10:83948 R, van Vliet SJ, van Duijnhoven GC,
54. Garrett WS, Chen LM, Kroschewski R, Middel J, Cornelissen IL, Nottet HS,
Ebersold M, Turley S, Trombetta S, KewalRamani VN, Littman DR, Figdor
Galan JE, Mellman I. 2000. Developmen- CG, van Kooyk Y. 2000. DC-SIGN, a den-
tal control of endocytosis in dendritic cells dritic cell-specific HIV-1-binding protein

by Cdc42. Cell 102:32534 that enhances trans-infection of T cells.
55. Marie JC, Kehren J, Trescol-Biemont Cell 100:58797
MC, Evlashev A, Valentin H, Walzer T, 63. Geijtenbeek TB, Torensma R, van Vliet
Tedone R, Loveland B, Nicolas JF, SJ, van Duijnhoven GC, Adema GJ,
Rabourdin-Combe C, Horvat B. 2001. van Kooyk Y, Figdor CG. 2000. Iden-
Mechanism of measles virus-induced sup- tification of DC-SIGN, a novel dendritic
pression of inflammatory immune re- cell-specific ICAM-3 receptor that sup-
sponses. Immunity 14:6979 ports primary immune responses. Cell
56. Werling D, Hope JC, Chaplin P, Collins 100:57585
RA, Taylor G, Howard CJ. 1999. Involve- 64. Medzhitov R, Janeway C Jr. 2000. Innate
ment of caveolae in the uptake of respira- immunity. N. Engl. J. Med. 343:33844
tory syncytial virus antigen by dendritic 65. Reis e Sousa C. 2001. Dendritic cells as
cells. J. Leuk. Biol. 66:5058 sensors of infection. Immunity 14:49598
57. Blank C, Fuchs H, Rappersberger K, 66. Aderem A, Ulevitch RJ. 2000. Toll-like
Rollinghoff M, Moll H. 1993. Parasi- receptors in the induction of the innate im-
tism of epidermal Langerhans cells in mune response. Nature 406:78287
experimental cutaneous leishmaniasis 67. Visintin A, Mazzoni A, Spitzer JH, Wyl-
with Leishmania major. J. Infect. Dis. lie DH, Dower SK, Segal DM. 2001.
167:41825 Regulation of Toll-like receptors in hu-
58. Gildea LA, Morris RE, Newman SL. man monocytes and dendritic cells. J. Im-
2001. Histoplasma capsulatum yeasts are munol. 166:24955
phagocytosed via very late antigen-5, 68. Muzio M, Bosisio D, Polentarutti N,
killed, and processed for antigen presenta- DAmico G, Stoppacciaro A, Mancinelli
tion by human dendritic cells. J. Immunol. R, vant Veer C, Penton-Rol G, Ruco LP,
166:104956 Allavena P, Mantovani A. 2000. Differen-
59. Cameron P, Pope M, Granelli-Piperno tial expression and regulation of toll-like
A, Steinman RM. 1996. Dendritic cells receptors (TLR) in human leukocytes: se-
and the replication of HIV-1. J. Leuk. Biol. lective expression of TLR3 in dendritic
59:15871 cells. J. Immunol. 164:59986004
60. Patterson S, Rae A, Hockey N, Gilmour 69. Michelsen KS, Aicher A, Mohaupt M,
J, Gotch F. 2001. Plasmacytoid dendritic Hartung T, Dimmeler S, Kirschning CJ,
cells are highly susceptible to human im- Schumann RR. 2001. The role of toll-like
munodeficiency virus type 1 infection and receptors (TLRs) in bacteria-induced mat-
release infectious virus. J. Virol. 75:6710 uration of murine dendritic cells (DCS).
13 Peptidoglycan and lipoteichoic acid are
inducers of dendritic cell maturation and dendritic cells through CD40 cross-
require TLR2. J. Biol. Chem. 276:25680 linking. J. Exp. Med. 180:126372
86 77. Rescigno M, Piguet V, Valzasina B,
70. Hertz C, Kiertscher S, Godowski P, Lens S, Zubler R, French L, Kindler V,
Bouis D, Norgard M, Roth M, Modlin Tschopp J, Ricciardi-Castagnoli P. 2000.
R. 2001. Microbial lipopeptides stimulate Fas engagement induces the maturation of
dendritic cell maturation via Toll-like re- dendritic cells (DCs), the release of inter-
ceptor 2. J. Immunol. 166:244450 leukin (IL)-1beta, and the production of
71. Nishiguchi M, Matsumoto M, Takao T, interferon gamma in the absence of IL-12
Hoshino M, Shimonishi Y, Tsuji S, Be- during DC-T cell cognate interaction: a
gum NA, Takeuchi O, Akira S, Toyosh- new role for Fas ligand in inflammatory
ima K, Seya T. 2001. Mycoplasma responses. J. Exp. Med. 192:166168
fermentans lipoprotein M161Ag-induced 78. Ohshima Y, Tanaka Y, Tozawa H, Taka-

cell activation is mediated by Toll-like re- hashi Y, Maliszewski C, Delespesse G.
ceptor 2: role of N-terminal hydrophobic 1997. Expression and function of OX40
portion in its multiple functions. J. Im- ligand on human dendritic cells. J. Im-
munol. 166:261016 munol. 159:383848
72. Jeannin P, Renno T, Goetsch L, Micon- 79. Regnault A, Lankar D, Lacabanne V,
net I, Aubry JP, Delneste Y, Herbault N, Rodriguez A, Thery C, Rescigno M,
Baussant T, Magistrelli G, Soulas C, Saito T, Verbeek S, Bonnerot C, Ricci-
Romero P, Cerottini JC, Bonnefoy JY. ardi-Castagnoli P, Amigorena S. 1999.
2000. OmpA targets dendritic cells, in- Fcgamma receptor-mediated induction
duces their maturation and delivers anti- of dendritic cell maturation and ma-
gen into the MHC class I presentation jor histocompatibility complex class
pathway. Nat. Immunol. 1:5029 I-restricted antigen presentation after im-
73. Underhill DM, Ozinsky A, Hajjar AM, mune complex internalization. J. Exp.
Stevens A, Wilson CB, Bassetti M, Med. 189:37180
Aderem A. 1999. The Toll-like receptor 80. Shi Y, Zheng W, Rock KL. 2000. Cell
2 is recruited to macrophage phagosomes injury releases endogenous adjuvants that
and discriminates between pathogens. Na- stimulate cytotoxic T cell responses. Proc.
ture 401:81115 Natl. Acad. Sci. USA 97:1459095
74. Kalinski P, Hilkens CM, Wierenga EA, 81. Rovere P, Vallinoto C, Bondanza A,
Kapsenberg ML. 1999. T-cell priming Crosti MC, Rescigno M, Ricciardi-Cas-
by type-1 and type-2 polarized dendritic tagnoli P, Rugarli C, Manfredi AA. 1998.
cells: the concept of a third signal. Im- Bystander apoptosis triggers dendritic cell
munol. Today 20:56167 maturation and antigen-presenting func-
75. Schuurhuis DH, Laban S, Toes RE, tion. J. Immunol. 161:446771
Ricciardi-Castagnoli P, Kleijmeer MJ, 82. Salio M, Cerundolo V, Lanzavecchia A.
van der Voort EI, Rea D, Offringa R, 2000. Dendritic cell maturation is in-
Geuze HJ, Melief CJ, Ossendorp F. 2000. duced by mycoplasma infection but not by
Immature dendritic cells acquire CD8(+) necrotic cells. Eur. J. Immunol. 30:7058
cytotoxic T lymphocyte priming capac- 83. Propato A, Cutrona G, Francavilla V,
ity upon activation by T helper cell- Ulivi M, Schiaffella E, Landt O, Dun-
independent or -dependent stimuli. J. Exp. bar R, Cerundolo V, Ferrarini M, Barn-
Med. 192:14550 aba V. 2001. Apoptotic cells overexpress
76. Caux C, Massacrier C, Vanbervliet B, vinculin and induce vinculin-specific cy-
Dubois B, Van Kooten C, Durand I, totoxic T-cell cross-priming. Nat. Med.
Banchereau J. 1994. Activation of human 7:80713
84. Basu S, Binder RJ, Suto R, Anderson regulated by different signaling pathways.
KM, Srivastava PK. 2000. Necrotic but J. Exp. Med. 188:217580
not apoptotic cell death releases heat 92. Ardeshna KM, Pizzey AR, Devereux S,
shock proteins, which deliver a partial Khwaja A. 2000. The PI3 kinase, p38 SAP
maturation signal to dendritic cells and kinase, and NF-kappaB signal transduc-
activate the NF-kappa B pathway. Int. Im- tion pathways are involved in the survival
munol. 12:153946 and maturation of lipopolysaccharide-
85. Singh-Jasuja H, Scherer HU, Hilf N, stimulated human monocyte-derived den-
Arnold-Schild D, Rammensee HG, Toes dritic cells. Blood 96:103946
RE, Schild H. 2000. The heat shock pro- 93. Vidalain PO, Azocar O, Servet-Delprat
tein gp96 induces maturation of dendritic C, Rabourdin-Combe C, Gerlier D,
cells and down-regulation of its receptor. Manie S. 2000. CD40 signaling in human
Eur. J. Immunol. 30:221115 dendritic cells is initiated within mem-

86. Vabulas RM, Ahmad-Nejad P, da Costa brane rafts. EMBO J. 19:330413
C, Miethke T, Kirschning CJ, Hacker H, 94. Aicher A, Shu GL, Magaletti D, Mulva-
Wagner H. 2001. Endocytosed heat shock nia T, Pezzutto A, Craxton A, Clark EA.
protein 60s use TLR2 and TLR4 to ac- 1999. Differential role for p38 mitogen-
tivate the toll/interleukin-1 receptor sig- activated protein kinase in regulating
naling pathway in innate immune cells. J. CD40-induced gene expression in den-
Biol. Chem. 276:31,33239 dritic cells and B cells. J. Immunol. 163:
87. Ohashi K, Burkart V, Flohe S, Kolb 578695
H. 2000. Cutting edge: heat shock protein 95. Lu B, Yu H, Chow C, Li B, Zheng W,
60 is a putative endogenous ligand of the Davis RJ, Flavell RA. 2001. GADD45
toll-like receptor-4 complex. J. Immunol. gamma mediates the activation of the p38
164:55861 and JNK MAP kinase pathways and cy-
88. Asea A, Kraeft SK, Kurt-Jones EA, tokine production in effector TH1 cells.
Stevenson MA, Chen LB, Finberg RW, Immunity 14:58390
Koo GC, Calderwood SK. 2000. HSP70 96. Lyakh LA, Koski GK, Telford W, Gress
stimulates cytokine production through a RE, Cohen PA, Rice NR. 2000. Bacterial
CD14-dependant pathway, demonstrating lipopolysaccharide, TNF-alpha, and cal-
its dual role as a chaperone and cytokine. cium ionophore under serum-free condi-
Nat. Med. 6:43542 tions promote rapid dendritic cell-like dif-
89. Kol A, Lichtman AH, Finberg RW, Libby ferentiation in CD14+ monocytes through
P, Kurt-Jones EA. 2000. Cutting edge: distinct pathways that activate NK-kappa
heat shock protein (HSP) 60 activates the B. J. Immunol. 165:364755
innate immune response: CD14 is an es- 97. Kaisho T, Akira S. 2001. Dendritic-
sential receptor for HSP60 activation of cell function in Toll-like receptor- and
mononuclear cells. J. Immunol. 164:13 MyD88-knockout mice. Trends Immunol.
17 22:7883
90. Geissmann F, Revy P, Regnault A, Lepel- 98. Muzio M, Natoli G, Saccani S, Lev-
letier Y, Dy M, Brousse N, Amigorena S, rero M, Mantovani A. 1998. The human
Hermine O, Durandy A. 1999. TGF-beta toll signaling pathway: divergence of nu-
1 prevents the noncognate maturation of clear factor kappaB and JNK/SAPK acti-
human dendritic Langerhans cells. J. Im- vation upstream of tumor necrosis factor
munol. 162:456775 receptor-associated factor 6 (TRAF6). J.
91. Rescigno M, Martino M, Sutherland CL, Exp. Med. 187:2097101
Gold MR, Ricciardi-Castagnoli P. 1998. 99. Kaisho T, Takeuchi O, Kawai T, Hoshino
Dendritic cell survival and maturation are K, Akira S. 2001. Endotoxin-induced
maturation of myd88-deficient dendritic following activation of CD34-derived hu-

cells. J. Immunol. 166:568894 man Langerhans cells. Proc. Natl. Acad.
100. Sallusto F, Lanzavecchia A. 2000. Under- Sci. USA 98:398287
standing dendritic cell and T-lymphocyte 109. Morel S, Levy F, Burlet-Schiltz O,
traffic through the analysis of chemokine Brasseur F, Probst-Kepper M, Peitrequin
receptor expression. Immunol. Rev. 177: AL, Monsarrat B, Van Velthoven R,
13440 Cerottini JC, Boon T, Gairin JE, Van den
101. Randolph GJ, Beaulieu S, Lebecque S, Eynde BJ. 2000. Processing of some anti-
Steinman RM, Muller WA. 1998. Differ- gens by the standard proteasome but not
entiation of monocytes into dendritic cells by the immunoproteasome results in poor
in a model of transendothelial trafficking. presentation by dendritic cells. Immunity
Science 282:48083 12:10717
102. Bevan MJ. 1976. Cross-priming for a sec- 110. Bates EE, Ravel O, Dieu MC, Ho S,
ondary cytotoxic response to minor H Guret C, Bridon JM, Ait-Yahia S, Briere
antigens with H-2 congenic cells which F, Caux C, Banchereau J, Lebecque S.
do not cross-react in the cytotoxic assay. 1997. Identification and analysis of a

J. Exp. Med. 143:128388 novel member of the ubiquitin family ex-
103. Cresswell P, Bangia N, Dick T, Diedrich pressed in dendritic cells and mature B
G. 1999. The nature of the MHC class I cells. Eur. J. Immunol. 27:247177
peptide loading complex. Immunol. Rev. 111. Norbury CC, Chambers BJ, Prescott AR,
172:2128 Ljunggren HG, Watts C. 1997. Consti-
104. Rescigno M, Citterio S, Thery C, Rittig tutive macropinocytosis allows TAP-dep-
M, Medaglini D, Pozzi G, Amigorena endent major histocompatibility complex
S, Ricciardi-Castagnoli P. 1998. Bacteria- class I presentation of exogenous soluble
induced neo-biosynthesis, stabilization, antigen by bone marrow-derived dendritic
and surface expression of functional class cells. Eur. J. Immunol. 27:28088
I molecules in mouse dendritic cells. Proc. 112. Kovacsovics-Bankowski M, Clark K, Be-
Natl. Acad. Sci. USA 95:522934 nacerraf B, Rock KL. 1993. Efficient
105. Cella M, Engering A, Pinet V, Pieters J, major histocompatibility complex class I
Lanzavecchia A. 1997. Inflammatory presentation of exogenous antigen upon
stimuli induce accumulation of MHC phagocytosis by macrophages. Proc. Natl.
class II complexes on dendritic cells. Na- Acad. Sci. USA 90:494246
ture 388:78287 113. Shen Z, Reznikoff G, Dranoff G, Rock
106. Cella M, Salio M, Sakakibara Y, Langen KL. 1997. Cloned dendritic cells can
H, Julkunen I, Lanzavecchia A. 1999. present exogenous antigens on both MHC
Maturation, activation, and protection class I and class II molecules. J. Immunol.
of dendritic cells induced by double- 158:272330
stranded RNA. J. Exp. Med. 189:82129 114. Reis e Sousa C, Germain RN. 1995. Ma-
107. Macagno A, Gilliet M, Sallusto F, Lanza- jor histocompatibility complex class I pre-
vecchia A, Nestle FO, Groettrup M. sentation of peptides derived from solu-
1999. Dendritic cells up-regulate im- ble exogenous antigen by a subset of cells
munoproteasomes and the proteasome engaged in phagocytosis. J. Exp. Med.
regulator PA28 during maturation. Eur. J. 182:84151
Immunol. 29:403742 115. Oh YK, Harding CV, Swanson JA. 1997.
108. MacAry PA, Lindsay M, Scott MA, The efficiency of antigen delivery from
Craig JI, Luzio JP, Lehner PJ. 2001. macrophage phagosomes into cytoplasm
Mobilization of MHC class I molecules for MHC class I-restricted antigen presen-
from late endosomes to the cell surface tation. Vaccine 15:51118
116. Wick MJ, Ljunggren HG. 1999. Process- with MHC class I. J. Immunol. Methods
ing of bacterial antigens for peptide pre- 248:18394
sentation on MHC class I molecules. Im- 124. Arnold D, Wahl C, Faath S, Rammensee
munol. Rev. 172:15362 HG, Schild H. 1997. Influences of trans-
117. Arrode G, Boccaccio C, Lule J, Allart porter associated with antigen processing
S, Moinard N, Abastado JP, Alam A, (TAP) on the repertoire of peptides as-
Davrinche C. 2000. Incoming human cy- sociated with the endoplasmic reticulum-
tomegalovirus pp65 (UL83) contained in resident stress protein gp96. J. Exp. Med.
apoptotic infected fibroblasts is cross- 186:46166
presented to CD8(+) T cells by dendritic 125. Binder RJ, Blachere NE, Srivastava PK.
cells. J. Virol. 74:1001824 2001. Heat shock protein-chaperoned
118. Nouri-Shirazi M, Banchereau J, Bell D, peptides but not free peptides introduced
Burkeholder S, Kraus ET, Davoust J, into the cytosol are presented efficiently
Palucka KA. 2000. Dendritic cells cap- by major histocompatibility complex I
ture killed tumor cells and present their molecules. J. Biol. Chem. 276:1716371
antigens to elicit tumor-specific immune 126. Singh-Jasuja H, Toes RE, Spee P, Munz
responses. J. Immunol. 165:3797803 C, Hilf N, Schoenberger SP, Ricciardi-
119. Fields RC, Shimizu K, Mule JJ. 1998. Castagnoli P, Neefjes J, Rammensee
Murine dendritic cells pulsed with whole HG, Arnold-Schild D, Schild H. 2000.
tumor lysates mediate potent antitumor Cross-presentation of glycoprotein 96-
immune responses in vitro and in vivo. associated antigens on major histocom-
Proc. Natl. Acad. Sci. USA 95:948287 patibility complex class I molecules re-
120. Wolfers J, Lozier A, Raposo G, Reg- quires receptor-mediated endocytosis. J.
nault A, Thery C, Masurier C, Flament C, Exp. Med. 191:196574
Pouzieux S, Faure F, Tursz T, Angevin E, 127. Haicheur N, Bismuth E, Bosset S, Ado-
Amigorena S, Zitvogel L. 2001. Tumor- tevi O, Warnier G, Lacabanne V, Reg-
derived exosomes are a source of shared nault A, Desaymard C, Amigorena S,
tumor rejection antigens for CTL cross- Ricciardi-Castagnoli P, Goud B, Fridman
priming. Nat. Med. 7:297303 WH, Johannes L, Tartour E. 2000. The
121. Machy P, Serre K, Leserman L. 2000. B subunit of Shiga toxin fused to a tumor
Class I-restricted presentation of exo- antigen elicits CTL and targets dendritic
genous antigen acquired by Fcgamma cells to allow MHC class I-restricted pre-
receptor-mediated endocytosis is regu- sentation of peptides derived from exoge-
lated in dendritic cells. Eur. J. Immunol. nous antigens. J. Immunol. 165:33018
30:84857 128. Guermonprez P, Khelef N, Blouin E,
122. Rovere P, Sabbadini MG, Vallinoto C, Rieu P, Ricciardi-Castagnoli P, Guiso N,
Fascio U, Rescigno M, Crosti M, Ricci- Ladant D, Leclerc C. 2001. The adeny-
ardi-Castagnoli P, Balestrieri G, Tincani late cyclase toxin of Bordetella pertus-
A, Manfredi AA. 1999. Dendritic cell sis binds to target cells via the alpha(M)
presentation of antigens from apoptotic beta(2) integrin (CD11b/CD18). J. Exp.
cells in a proinflammatory context: role of Med. 193:103544
opsonizing anti-beta2-glycoprotein I anti- 129. Yewdell JW, Norbury CC, Bennink JR.
bodies. Arthritis Rheumatol. 42:141220 1999. Mechanisms of exogenous antigen
123. Wallace PK, Tsang KY, Goldstein J, Cor- presentation by MHC class I molecules in
reale P, Jarry TM, Schlom J, Guyre PM, vitro and in vivo: implications for gener-
Ernstoff MS, Fanger MW. 2001. Exo- ating CD8+ T cell responses to infectious
genous antigen targeted to FcgammaRI on agents, tumors, transplants, and vaccines.
myeloid cells is presented in association Adv. Immunol. 73:177
130. Gromme M, Uytdehaag FG, Janssen H, endothelial cells to CD8+ T cells results
Calafat J, van Binnendijk RS, Kenter MJ, in antigen-specific T-cell tolerance. Nat.
Tulp A, Verwoerd D, Neefjes J. 1999. Re- Med. 6:134854
cycling MHC class I molecules and endo- 139. Huang AY, Bruce AT, Pardoll DM, Lev-
somal peptide loading. Proc. Natl. Acad. itsky HI. 1996. In vivo cross-priming of
Sci. USA 96:1032631 MHC class I-restricted antigens requires
131. Kleijmeer MJ, Escola JM, UytdeHaag the TAP transporter. Immunity 4:34955
FG, Jakobson E, Griffith JM, Osterhaus 140. Sigal LJ, Crotty S, Andino R, Rock KL.
AD, Stoorvogel W, Melief CJ, Rabouille 1999. Cytotoxic T-cell immunity to virus-
C, Geuze HJ. 2001. Antigen loading of infected non-haematopoietic cells re-
MHC class I molecules in the endocytic quires presentation of exogenous antigen.
tract. Traffic 2:12437 Nature 398:7780
132. Guyre CA, Barreda ME, Swink SL, 141. Cresswell P. 1996. Invariant chain struc-
Fanger MW. 2001. Colocalization of Fc ture and MHC class II function. Cell
gamma RI-targeted antigen with class I 84:5057
MHC: implications for antigen process- 142. Villadangos JA, Bryant RA, Deussing J,
ing. J. Immunol. 166:246978 Driessen C, Lennon-Dumenil AM, Riese
133. Kovacsovics-Bankowski M, Rock KL. RJ, Roth W, Saftig P, Shi GP, Chapman
1995. A phagosome-to-cytosol pathway HA, Peters C, Ploegh HL. 1999. Pro-
for exogenous antigens presented on teases involved in MHC class II antigen
MHC class I molecules. Science 267:243 presentation. Immunol. Rev. 172:10920
46 143. Kropshofer H, Hammerling GJ, Vogt AB.
134. Garbi N, Tan P, Diehl AD, Chambers BJ, 1999. The impact of the non-classical
Ljunggren HG, Momburg F, Hammerling MHC proteins HLA-DM and HLA-DO
GJ. 2000. Impaired immune responses on loading of MHC class II molecules.
and altered peptide repertoire in tapasin- Immunol. Rev. 172:26778
deficient mice. Nat. Immunol. 1:23438 144. Watts C. 2001. Antigen processing in the
135. Norbury CC, Hewlett LJ, Prescott AR, endocytic compartment. Curr. Opin. Im-
Shastri N, Watts C. 1995. Class I MHC munol. 13:2631
presentation of exogenous soluble anti- 145. Inaba K, Turley S, Iyoda T, Yamaide F,
gen via macropinocytosis in bone marrow Shimoyama S, Reis e Sousa C, Germain
macrophages. Immunity 3:78391 RN, Mellman I, Steinman RM. 2000.
136. Rodriguez A, Regnault A, Kleijmeer M, The formation of immunogenic major his-
Ricciardi-Castagnoli P, Amigorena S. tocompatibility complex class II-peptide
1999. Selective transport of internalized ligands in lysosomal compartments of
antigens to the cytosol for MHC class I dendritic cells is regulated by inflam-
presentation in dendritic cells. Nat. Cell matory stimuli. J. Exp. Med. 191:927
Biol. 1:36268 36
137. Merkenschlager M, Power MO, Pircher 146. Fiebiger E, Meraner P, Weber E, Fang
H, Fisher AG. 1999. Intrathymic dele- IF, Stingl G, Ploegh H, Maurer D. 2001.
tion of MHC class I-restricted cytotoxic Cytokines regulate proteolysis in major
T cell precursors by constitutive cross- histocompatibility complex class II-de-
presentation of exogenous antigen. Eur. J. pendent antigen presentation by dendritic
Immunol. 29:147786 cells. J. Exp. Med. 193:88192
138. Limmer A, Ohl J, Kurts C, Ljunggren 147. Bevec T, Stoka V, Pungercic G, Dolenc
HG, Reiss Y, Groettrup M, Momburg F, I, Turk V. 1996. Major histocompatibil-
Arnold B, Knolle PA. 2000. Efficient pre- ity complex class II-associated p41 invari-
sentation of exogenous antigen by liver ant chain fragment is a strong inhibitor
of lysosomal cathepsin L. J. Exp. Med. pendently of invariant chain degradation.

183:133138 Immunity 14:73949
148. Pierre P, Shachar I, Matza D, Gatti E, 156. Santambrogio L, Sato AK, Carven GJ,
Flavell RA, Mellman I. 2000. Invariant Belyanskaya SL, Strominger JL, Stern
chain controls H2-M proteolysis in mouse LJ. 1999. Extracellular antigen process-
splenocytes and dendritic cells. J. Exp. ing and presentation by immature den-
Med. 191:105762 dritic cells. Proc. Natl. Acad. Sci. USA
149. Pierre P, Turley SJ, Gatti E, Hull M, 96:1505661
Meltzer J, Mirza A, Inaba K, Steinman 157. Askew D, Chu RS, Krieg AM, Hard-
RM, Mellman I. 1997. Developmental ing CV. 2000. CpG DNA induces mat-
regulation of MHC class II transport in uration of dendritic cells with distinct
mouse dendritic cells. Nature 388:78792 effects on nascent and recycling MHC-
150. Pierre P, Mellman I. 1998. Developmen- II antigen-processing mechanisms. J. Im-

tal regulation of invariant chain prote- munol. 165:688995
olysis controls MHC class II trafficking 158. Turley SJ, Inaba K, Garrett WS, Ebersold
in mouse dendritic cells. Cell 93:1135 M, Unternaehrer J, Steinman RM, Mell-

45 man I. 2000. Transport of peptide-MHC
151. Nakagawa TY, Brissette WH, Lira PD, class II complexes in developing dendritic
Griffiths RJ, Petrushova N, Stock J, Mc- cells. Science 288:52227
Neish JD, Eastman SE, Howard ED, 159. Baron C, Raposo G, Scholl SM, Bau-
Clarke SR, Rosloniec EF, Elliott EA, singer H, Tenza D, Bohbot A, Pouil-
Rudensky AY. 1999. Impaired invariant lart P, Goud B, Hanau D, Salamero J.
chain degradation and antigen presen- 2001. Modulation of MHC class II
tation and diminished collagen-induced transport and lysosome distribution by
arthritis in cathepsin S null mice. Immu- macrophage-colony stimulating factor in
nity 10:20717 human dendritic cells derived from mono-
152. Shi GP, Villadangos JA, Dranoff G, cytes. J. Cell Sci. 114:9991010
Small C, Gu L, Haley KJ, Riese R, 160. Porcelli SA, Modlin RL. 1999. The CD1
Ploegh HL, Chapman HA. 1999. Cathep- system: antigen-presenting molecules for
sin S required for normal MHC class II T cell recognition of lipids and glycol-
peptide loading and germinal center de- ipids. Annu. Rev. Immunol. 17:297329
velopment. Immunity 10:197206 161. Matsuda JL, Kronenberg M. 2001. Pre-
153. Driessen C, Bryant RA, Lennon-Dume- sentation of self and microbial lipids by
nil AM, Villadangos JA, Bryant PW, Shi CD1 molecules. Curr. Opin. Immunol.
GP, Chapman HA, Ploegh HL. 1999. 13:1925
Cathepsin S controls the trafficking and 162. Jayawardena-Wolf J, Bendelac A. 2001.
maturation of MHC class II molecules in CD1 and lipid antigens: intracellular path-
dendritic cells. J. Cell Biol. 147:77590 ways for antigen presentation. Curr. Opin.
154. Rovere P, Zimmermann VS, Forquet F, Immunol. 13:10913
Demandolx D, Trucy J, Ricciardi-Casta- 163. Sugita M, Grant EP, van Donselaar E,
gnoli P, Davoust J. 1998. Dendritic cell Hsu VW, Rogers RA, Peters PJ, Brenner
maturation and antigen presentation in the MB. 1999. Separate pathways for antigen
absence of invariant chain. Proc. Natl. presentation by CD1 molecules. Immunity
Acad. Sci. USA 95:106772 11:74352
155. Villadangos JA, Cardoso M, Steptoe RJ, 164. Salamero J, Bausinger H, Mommaas
van Berkel D, Pooley J, Carbone FR, AM, Lipsker D, Proamer F, Cazenave
Shortman K. 2001. MHC class II expres- JP, Goud B, de la Salle H, Hanau D.
sion is regulated in dendritic cells inde- 2001. CD1a molecules traffic through the
early recycling endosomal pathway in hu- presenting function of the mouse CD1
man Langerhans cells. J. Invest. Derma- molecule. Ann. N.Y. Acad. Sci. 778:288
tol. 116:4018 96
165. Sugita M, van Der Wel N, Rogers RA, 173. Yang OO, Racke FK, Nguyen PT, Gau-
Peters PJ, Brenner MB. 2000. CD1c sling R, Severino ME, Horton HF, Byrne
molecules broadly survey the endocytic MC, Strominger JL, Wilson SB. 2000.
system. Proc. Natl. Acad. Sci. USA 97: CD1d on myeloid dendritic cells stimu-
844550 lates cytokine secretion from and cytolytic
166. Schaible UE, Hagens K, Fischer K, Col- activity of V alpha 24J alpha Q T cells: a
lins HL, Kaufmann SH. 2000. Intersec- feedback mechanism for immune regula-
tion of group I CD1 molecules and tion. J. Immunol. 165:375662
mycobacteria in different intracellular 174. Kitamura H, Iwakabe K, Yahata T, Nishi-
compartments of dendritic cells. J. Immu- mura S, Ohta A, Ohmi Y, Sato M, Takeda

nol. 164:484352 K, Okumura K, Van Kaer L, Kawano T,
167. Shamshiev A, Donda A, Prigozy TI, Taniguchi M, Nishimura T. 1999. The
Mori L, Chigorno V, Benedict CA, Kap- natural killer T (NKT) cell ligand alpha-
pos L, Sonnino S, Kronenberg M, De galactosylceramide demonstrates its im-
Libero G. 2000. The alphabeta T cell re- munopotentiating effect by inducing in-
sponse to self-glycolipids shows a novel terleukin (IL)-12 production by dendritic
mechanism of CD1b loading and a re- cells and IL-12 receptor expression on
quirement for complex oligosaccharides. NKT cells. J. Exp. Med. 189:112128
Immunity 13:25564 175. Nishimura T, Kitamura H, Iwakabe K,
168. Briken V, Jackman RM, Watts GF, Yahata T, Ohta A, Sato M, Takeda K,
Rogers RA, Porcelli SA. 2000. Human Okumura K, Van Kaer L, Kawano T,
CD1b and CD1c isoforms survey different Taniguchi M, Nakui M, Sekimoto M,
intracellular compartments for the presen- Koda T. 2000. The interface between in-
tation of microbial lipid antigens. J. Exp. nate and acquired immunity: glycolipid
Med. 192:28188 antigen presentation by CD1d-expressing
169. Park SH, Weiss A, Benlagha K, Kyin T, dendritic cells to NKT cells induces the
Teyton L, Bendelac A. 2001. The mouse differentiation of antigen-specific cyto-
CD1d-restricted repertoire is dominated toxic T lymphocytes. Int. Immunol. 12:
by a few autoreactive T cell receptor fam- 98794
ilies. J. Exp. Med. 193:893904 176. Ikarashi Y, Mikami R, Bendelac A,
170. Gumperz JE, Roy C, Makowska A, Lum Terme M, Chaput N, Tursz T, Angevin E,
D, Sugita M, Podrebarac T, Koezuka Y, Lemonnier FA, Wakasugi H, Zitvogel L.
Porcelli SA, Cardell S, Brenner MB, Be- 2001. Dendritic cell maturation overrules
har SM. 2000. Murine CD1d-restricted T H-2D-mediated natural killer (NKT) cell
cell recognition of cellular lipids. Immu- inhibition: critical role for B7 in CD1d-
nity 12:21121 dependent NKT cell IFN production. J.
171. Chiu YH, Jayawardena J, Weiss A, Lee Exp. Med. 194:117986
D, Park SH, Dautry-Varsat A, Bendelac 177. Angenieux C, Salamero J, Fricker D,
A. 1999. Distinct subsets of CD1d- Cazenave JP, Goud B, Hanau D, de La
restricted T cells recognize self-antigens Salle H. 2000. Characterization of CD1e,
loaded in different cellular compartments. a third type of CD1 molecule expressed in
J. Exp. Med. 189:10310 dendritic cells. J. Biol. Chem. 275:37757
172. Tangri S, Holcombe HR, Castano AR, 64
Miller JE, Teitell M, Huse WE, Peter- 178. Inaba K, Metlay JP, Crowley MT, Stein-
son PA, Kronenberg M. 1996. Antigen- man RM. 1990. Dendritic cells pulsed
with protein antigens in vitro can prime 186. Casares S, Inaba K, Brumeanu TD,
antigen-specific, MHC-restricted T cells Steinman RM, Bona CA. 1997. Anti-
in situ [published erratum appears in J. gen presentation by dendritic cells af-
Exp. Med. [1990 Oct 1; 172(4):1275]. J. ter immunization with DNA encoding a
Exp. Med. 172:63140 major histocompatibility complex class
179. Mayordomo JI, Zorina T, Storkus WJ, II-restricted viral epitope. J. Exp. Med.
Zitvogel L, Celluzzi C, Falo LD, Melief 186:148186
CJ, Ildstad ST, Kast WM, Deleo AB, 187. Porgador A, Irvine KR, Iwasaki A, Bar-
et al. 1995. Bone marrow-derived den- ber BH, Restifo NP, Germain RN. 1998.
dritic cells pulsed with synthetic tumour Predominant role for directly transfected
peptides elicit protective and therapeutic dendritic cells in antigen presentation to
antitumour immunity. Nat. Med. 1:1297 CD8+ T cells after gene gun immuniza-
302 tion. J. Exp. Med. 188:107582

180. Moll H, Fuchs H, Blank C, Rollinghoff 188. Ingulli E, Mondino A, Khoruts A, Jenk-
M. 1993. Langerhans cells transport ins MK. 1997. In vivo detection of den-
Leishmania major from the infected skin dritic cell antigen presentation to CD4(+)
to the draining lymph node for presenta- T cells. J. Exp. Med. 185:213341
tion to antigen-specific T cells. Eur. J. Im- 189. Ridge JP, Fuchs EJ, Matzinger P. 1996.
munol. 23:1595601 Neonatal tolerance revisited: turning on
181. Usherwood EJ, Hogg TL, Woodland DL. newborn T cells with dendritic cells. Sci-
1999. Enumeration of antigen-presenting ence 271:172326
cells in mice infected with Sendai virus. 190. Pulendran B, Smith JL, Jenkins M, Scho-
J. Immunol. 162:335055 enborn M, Maraskovsky E, Maliszewski
182. Bujdoso R, Hopkins J, Dutia BM, Young CR. 1998. Prevention of peripheral tol-
P, McConnell I. 1989. Characterization of erance by a dendritic cell growth factor:
sheep afferent lymph dendritic cells and flt3 ligand as an adjuvant. J. Exp. Med.
their role in antigen carriage. J. Exp. Med. 188:207582
170:1285301 191. Steptoe RJ, Fu F, Li W, Drakes ML, Lu
183. Guery JC, Adorini L. 1995. Dendritic L, Demetris AJ, Qian S, McKenna HJ,
cells are the most efficient in present- Thomson AW. 1997. Augmentation of
ing endogenous naturally processed self- dendritic cells in murine organ donors
epitopes to class II-restricted T cells. J. by Flt3 ligand alters the balance between
Immunol. 154:53644 transplant tolerance and immunity. J. Im-
184. Zhong G, Sousa CR, Germain RN. munol. 159:548391
1997. Antigen-unspecific B cells and lym- 192. Mayordomo JI, Loftus DJ, Sakamoto H,
phoid dendritic cells both show extensive De Cesare CM, Appasamy PM, Lotze
surface expression of processed antigen- MT, Storkus WJ, Appella E, DeLeo AB.
major histocompatibility complex class II 1996. Therapy of murine tumors with p53
complexes after soluble protein exposure wild-type and mutant sequence peptide-
in vivo or in vitro. J. Exp. Med. 186:673 based vaccines. J. Exp. Med. 183:1357
82 65
185. Reis e Sousa C, Germain RN. 1999. 193. Ludewig B, Odermatt B, Landmann S,
Analysis of adjuvant function by direct Hengartner H, Zinkernagel RM. 1998.
visualization of antigen presentation in Dendritic cells induce autoimmune dia-
vivo: endotoxin promotes accumulation betes and maintain disease via de novo
of antigen-bearing dendritic cells in the T formation of local lymphoid tissue. J. Exp.
cell areas of lymphoid tissue. J. Immunol. Med. 188:1493501
162:655261 194. Schreurs MW, Eggert AA, de Boer AJ,
Vissers JL, van Hall T, Offringa R, Fig- tolerogenic or immunogenic fashion. J.

dor CG, Adema GJ. 2000. Dendritic Immunol. 157:140614
cells break tolerance and induce pro- 201a. Hawiger D, Inaba K, Dorsett Y, Guo
tective immunity against a melanocyte M, Mahnke K, Rivera M, Ravetch JV,
differentiation antigen in an autologous Steinman RM, Nussenzweig MC. 2001.
melanoma model. Cancer Res. 60:6995 Dendritic cells induce peripheral T cell
7001 unresponsiveness under steady state con-
195. Ludewig B, Ochsenbein AF, Odermatt ditions in vivo. J. Exp. Med. 194:76979
B, Paulin D, Hengartner H, Zinkernagel 202. Thomson AW, Lu L. 1999. Are dendritic
RM. 2000. Immunotherapy with den- cells the key to liver transplant tolerance?
dritic cells directed against tumor anti- Immunol. Today 20:2732
gens shared with normal host cells re- 203. Huang AY, Golumbek P, Ahmadzadeh
sults in severe autoimmune disease. J. M, Jaffee E, Pardoll D, Levitsky H.

Exp. Med. 191:795804 1994. Role of bone marrow-derived cells
196. Brocker T, Riedinger M, Karjalainen K. in presenting MHC class I-restricted tu-
1997. Targeted expression of major his- mor antigens. Science 264:96165

tocompatibility complex (MHC) class 204. Lenz LL, Butz EA, Bevan MJ. 2000.
II molecules demonstrates that dendritic Requirements for bone marrow-derived
cells can induce negative but not positive antigen-presenting cells in priming cy-
selection of thymocytes in vivo. J. Exp. totoxic T cell responses to intracellular
Med. 185:54150 pathogens. J. Exp. Med. 192:113542
197. Kyewski BA, Fathman CG, Rouse RV. 205. Sigal LJ, Rock KL. 2000. Bone marrow-
1986. Intrathymic presentation of circu- derived antigen-presenting cells are re-
lating non-MHC antigens by medullary quired for the generation of cytotoxic
dendritic cells. An antigen-dependent T lymphocyte responses to viruses and
microenvironment for T cell differenti- use transporter associated with anti-
ation. J. Exp. Med. 163:23146 gen presentation (TAP)-dependent and
198. Crowley M, Inaba K, Steinman RM. -independent pathways of antigen pre-
1990. Dendritic cells are the principal sentation. J. Exp. Med. 192:114350
cells in mouse spleen bearing immuno- 206. Miller JF, Kurts C, Allison J, Kosaka H,
genic fragments of foreign proteins. J. Carbone F, Heath WR. 1998. Induction
Exp. Med. 172:38386 of peripheral CD8+ T-cell tolerance by
199. Guery JC, Ria F, Galbiati F, Smiroldo cross-presentation of self antigens. Im-
S, Adorini L. 1997. The mode of protein munol. Rev. 165:26777
antigen administration determines pref- 207. Kurts C, Cannarile M, Klebba I, Broc-
erential presentation of peptide-class II ker T. 2001. Dendritic cells are sufficient
complexes by lymph node dendritic or B to cross-present self-antigens to CD8 T
cells. Int. Immunol. 9:915 cells in vivo. J. Immunol. 166:143942
200. Inaba K, Pack M, Inaba M, Sakuta H, 208. Chiodoni C, Paglia P, Stoppacciaro A,
Isdell F, Steinman RM. 1997. High lev- Rodolfo M, Parenza M, Colombo MP.
els of a major histocompatibility com- 1999. Dendritic cells infiltrating tu-
plex II-self peptide complex on den- mors cotransduced with granulocyte/
dritic cells from the T cell areas of macrophage colony-stimulating factor
lymph nodes. J. Exp. Med. 186:665 (GM-CSF) and CD40 ligand genes
72 take up and present endogenous tumor-
201. Finkelman FD, Lees A, Birnbaum R, associated antigens, and prime naive
Gause WC, Morris SC. 1996. Dendritic mice for a cytotoxic T lymphocyte re-
cells can present antigen in vivo in a sponse. J. Exp. Med. 190:12533
209. Li M, Davey GM, Sutherland RM, cells generate an immune response after
Kurts C, Lew AM, Hirst C, Carbone FR, subcutaneous injection without homing
Heath WR. 2001. Cell-associated oval- to the draining lymph node. J. Exp. Med.
bumin is cross-presented much more ef- 189:59398
ficiently than soluble ovalbumin in vivo. 219. Liu YJ, Kanzler H, Soumelis V, Gilliet
J. Immunol. 166:6099103 M. 2001. Dendritic cell lineage, plastic-
210. Banchereau J, Pulendran B, Steinman ity and cross-regulation. Nat. Immunol.
R, Palucka K. 2000. Will the making of 2:58589
plasmacytoid dendritic cells in vitro help 220. Grouard G, Rissoan MC, Filgueira L,
unravel their mysteries? J. Exp. Med. Durand I, Banchereau J, Liu YJ. 1997.
192:F3944 The enigmatic plasmacytoid T cells de-
211. Shortman K, Wu L. 2001. Parentage velop into dendritic cells with interleukin
and heritage of dendritic cells. Blood (IL)-3 and CD40-ligand. J. Exp. Med.
97:3325 185:110111
212. Ruedl C, Bachmann MF. 1999. CTL 221. Rissoan MC, Soumelis V, Kadowaki N,
priming by CD8(+) and CD8() den- Grouard G, Briere F, de Waal Malefyt

dritic cells in vivo. Eur. J. Immunol. 29: R, Liu YJ. 1999. Reciprocal control of
376267 T helper cell and dendritic cell differen-
213. Pooley JL, Heath WR, Shortman K. tiation. Science 283:118386
2001. Cutting edge: intravenous solu- 222. Freeman GJ, Long AJ, Iwai Y, Bourque
ble antigen is presented to cd4 t cells K, Chernova T, Nishimura H, Fitz LJ,
by CD8() dendritic cells, but cross- Malenkovich N, Okazaki T, Byrne MC,
presented to CD8 T cells by CD8(+) den- Horton HF, Fouser L, Carter L, Ling
dritic cells. J. Immunol. 166:532730 V, Bowman MR, Carreno BM, Collins
214. den Haan JM, Lehar SM, Bevan MJ. M, Wood CR, Honjo T. 2000. Engage-
2000. CD8(+) but not CD8() dendritic ment of the PD-1 immunoinhibitory re-
cells cross-prime cytotoxic T cells in ceptor by a novel B7 family member
vivo. J. Exp. Med. 192:168596 leads to negative regulation of lympho-
215. Maldonado-Lopez R, De Smedt T, cyte activation. J. Exp. Med. 192:1027
Michel P, Godfroid J, Pajak B, Heirman 34
C, Thielemans K, Leo O, Urbain J, 223. Dong H, Zhu G, Tamada K, Chen L.
Moser M. 1999. CD8alpha+ and 1999. B7-H1, a third member of the B7
CD8alpha subclasses of dendritic family, co-stimulates T-cell proliferation
cells direct the development of distinct and interleukin-10 secretion. Nat. Med.
T helper cells in vivo. J. Exp. Med. 189: 5:136569
58792 224. Tseng SY, Otsuji M, Gorski K, Huang
216. Pulendran B, Smith JL, Caspary G, Bra- X, Slansky JE, Pai SI, Shalabi A,
sel K, Pettit D, Maraskovsky E, Mal- Shin T, Pardoll DM, Tsuchiya H. 2001.
iszewski CR. 1999. Distinct dendritic B7-DC, a new dendritic cell molecule
cell subsets differentially regulate the with potent costimulatory properties for
class of immune response in vivo. Proc. T cells. J. Exp. Med. 193:83946
Natl. Acad. Sci. USA 96:103641 225. Shuford WW, Klussman K, Tritchler
217. Suss G, Shortman K. 1996. A sub- DD, Loo DT, Chalupny J, Siadak AW,
class of dendritic cells kills CD4 T cells Brown TJ, Emswiler J, Raecho H,
via Fas/Fas-ligand- induced apoptosis. J. Larsen CP, Pearson TC, Ledbetter JA,
Exp. Med. 183:178996 Aruffo A, Mittler RS. 1997. 4-1BB co-
218. Smith AL, de St Groth BF. 1999. stimulatory signals preferentially induce
Antigen-pulsed CD8alpha+ dendritic CD8+ T cell proliferation and lead to the
amplification in vivo of cytotoxic T cell peptide-induced peripheral cytotoxic T-

responses. J. Exp. Med. 186:4755 lymphocyte tolerance and augments
226. Lane P. 2000. Role of OX40 signals in anti-tumor vaccine efficacy. Nat. Med.
coordinating CD4 T cell selection, mi- 5:77479
gration, and cytokine differentiation in T 235. Lu Z, Yuan L, Zhou X, Sotomayor E,
helper (Th)1 and Th2 cells. J. Exp. Med. Levitsky HI, Pardoll DM. 2000. CD40-
191:2016 independent pathways of T cell help for
227. Ruedl C, Kopf M, Bachmann MF. 1999. priming of CD8(+) cytotoxic T lympho-
CD8(+) T cells mediate CD40-inde- cytes. J. Exp. Med. 191:54150
pendent maturation of dendritic cells in 236. Hirao M, Onai N, Hiroishi K, Watkins
vivo. J. Exp. Med. 189:187584 SC, Matsushima K, Robbins PD, Lotze
228. Bennett SR, Carbone FR, Karamalis F, MT, Tahara H. 2000. CC chemokine
Miller JF, Heath WR. 1997. Induction receptor-7 on dendritic cells is induced
of a CD8+ cytotoxic T lymphocyte re- after interaction with apoptotic tumor
sponse by cross-priming requires cog- cells: critical role in migration from the
nate CD4+ T cell help. J. Exp. Med. tumor site to draining lymph nodes. Can-
186:6570 cer Res. 60:220917
229. Kurts C, Carbone FR, Barnden M, 237. Steinman RM, Turley S, Mellman I,
Blanas E, Allison J, Heath WR, Miller Inaba K. 2000. The induction of toler-
JF. 1997. CD4+ T cell help impairs CD8+ ance by dendritic cells that have captured
T cell deletion induced by cross-presen- apoptotic cells. J. Exp. Med. 191:411
tation of self-antigens and favors autoim- 16
munity. J. Exp. Med. 186:205762 238. Roncarolo MG, Levings MK, Traversari
230. Bennett SR, Carbone FR, Karamalis F, C. 2001. Differentiation of T regulatory
Flavell RA, Miller JF, Heath WR. 1998. cells by immature dendritic cells. J. Exp.
Help for cytotoxic-T-cell responses is Med. 193:F59
mediated by CD40 signalling. Nature 239. Jonuleit H, Schmitt E, Steinbrink K,
393:47880 Enk AH. 2001. Dendritic cells as a tool
231. Ridge JP, Di Rosa F, Matzinger P. to induce anergic and regulatory T cells.
1998. A conditioned dendritic cell can Trends Immunol. 22:394400
be a temporal bridge between a CD4+ 240. Jonuleit H, Schmitt E, Schuler G,
T-helper and a T-killer cell. Nature 393: Knop J, Enk AH. 2000. Induction of
47478 interleukin 10-producing, nonprolifera-
232. Schoenberger SP, Toes RE, van der ting CD4(+) T cells with regulatory prop-
Voort EI, Offringa R, Melief CJ. 1998. erties by repetitive stimulation with allo-
T-cell help for cytotoxic T lymphocytes geneic immature human dendritic cells.
is mediated by CD40-CD40L interac- J. Exp. Med. 192:121322
tions. Nature 393:48083 241. Dhodapkar MV, Steinman RM, Kraso-
233. Mackey MF, Gunn JR, Maliszewsky vsky J, Munz C, Bhardwaj N. 2001.
C, Kikutani H, Noelle RJ, Barth RJ Jr. Antigen-specific inhibition of effector T
1998. Dendritic cells require maturation cell function in humans after injection
via CD40 to generate protective antitu- of immature dendritic cells. J. Exp. Med.
mor immunity. J. Immunol. 161:2094 193:23338
98 242. Fu F, Li Y, Qian S, Lu L, Chambers F,
234. Diehl L, den Boer AT, Schoenberger SP, Starzl TE, Fung JJ, Thomson AW.
van der Voort EI, Schumacher TN, 1996. Costimulatory molecule-deficient
Melief CJ, Offringa R, Toes RE. 1999. dendritic cell progenitors (MHC
CD40 activation in vivo overcomes class II+, CD80dim, CD86) prolong
cardiac allograft survival in nonimmuno- monocyte-derived dendritic cells induce

suppressed recipients. Transplantation naive T cell differentiation into T helper
62:65965 cell type 2 (Th2) or Th1/Th2 effectors.
243. Cella M, Scheidegger D, Palmer-Leh- Role of stimulator/responder ratio. J.
mann K, Lane P, Lanzavecchia A, Alber Exp. Med. 192:40512
G. 1996. Ligation of CD40 on dendritic 252. Langenkamp A, Messi M, Lanzavec-
cells triggers production of high levels of chia A, Sallusto F. 2000. Kinetics of den-
interleukin-12 and enhances T cell stim- dritic cell activation: impact on priming
ulatory capacity: T-T help via APC acti- of TH1, TH2 and nonpolarized T cells.
vation. J. Exp. Med. 184:74752 Nat. Immunol. 1:31116
244. Hartmann G, Weiner GJ, Krieg AM. 253. Iezzi G, Karjalainen K, Lanzavecchia A.
1999. CpG DNA: a potent signal for 1998. The duration of antigenic stimu-
growth, activation, and maturation of hu- lation determines the fate of naive and
man dendritic cells. Proc. Natl. Acad. effector T cells. Immunity 8:8995
Sci. USA 96:930510 254. Bevan MJ, Fink PJ. 2001. The CD8 re-
245. Cella M, Jarrossay D, Facchetti F, Ale- sponse on autopilot. Nat. Immunol. 2:

bardi O, Nakajima H, Lanzavecchia A, 38182
Colonna M. 1999. Plasmacytoid mono- 255. Josien BR, Li HL, Ingulli E, Sarma S,
cytes migrate to inflamed lymph nodes Wong BR, Vologodskaia M, Steinman
and produce large amounts of type I in- RM, Choi Y. 2000. TRANCE, a tumor
terferon. Nat. Med. 5:91923 necrosis factor family member, enhances
246. Cella M, Facchetti F, Lanzavecchia A, the longevity and adjuvant properties of
Colonna M. 2000. Plasmacytoid den- dendritic cells in vivo. J. Exp. Med. 191:
dritic cells activated by influenza virus 495502
and CD40L drive a potent TH1 polariza- 256. De Smedt T, Pajak B, Klaus GG, Noelle
tion. Nat. Immunol. 1:30510 RJ, Urbain J, Leo O, Moser M. 1998.
247. Kadowaki N, Antonenko S, Lau JY, Liu Antigen-specific T lymphocytes regu-
YJ. 2000. Natural interferon alpha/beta- late lipopolysaccharide-induced apopto-
producing cells link innate and adaptive sis of dendritic cells in vivo. J. Immunol.
immunity. J. Exp. Med. 192:21926 161:447679
248. Pulendran B, Palucka K, Banchereau 257. Hermans IF, Ritchie DS, Yang J,
J. 2001. Sensing pathogens and tuning Roberts JM, Ronchese F. 2000. CD8+ T
immune responses. Science 293:25356 cell-dependent elimination of dendritic
249. Iwasaki A, Kelsall BL. 1999. Freshly cells in vivo limits the induction of anti-
isolated Peyers patch, but not spleen, tumor immunity. J. Immunol. 164:3095
dendritic cells produce interleukin 10 101
and induce the differentiation of T helper 258. Ludewig B, Bonilla WV, Dumrese T,
type 2 cells. J. Exp. Med. 190:22939 Odermatt B, Zinkernagel RM, Hengart-
250. Stumbles PA, Thomas JA, Pimm CL, ner H. 2001. Perforin-independent regu-
Lee PT, Venaille TJ, Proksch S, Holt lation of dendritic cell homeostasis by
PG. 1998. Resting respiratory tract den- CD8(+) T cells in vivo: implications for
dritic cells preferentially stimulate T adaptive immunotherapy. Eur. J. Immu-
helper cell type 2 (Th2) responses and nol. 31:177279
require obligatory cytokine signals for 259. Ludewig B, Oehen S, Barchiesi F,
induction of Th1 immunity. J. Exp. Med. Schwendener RA, Hengartner H, Zin-
188:201931 kernagel RM. 1999. Protective antiviral
251. Tanaka H, Demeure CE, Rubio M, De- cytotoxic T cell memory is most effi-
lespesse G, Sarfati M. 2000. Human ciently maintained by restimulation via
dendritic cells. J. Immunol. 163:1839 to apoptosis through the Fas (CD95/

44 APO-1) pathway. J. Immunol. 163:
260. McLellan A, Heldmann M, Terbeck G, 530311
Weih F, Linden C, Brocker EB, Lever- 267. Leverkus M, Walczak H, McLellan A,
kus M, Kampgen E. 2000. MHC class Fries HW, Terbeck G, Brocker EB,
II and CD40 play opposing roles in Kampgen E. 2000. Maturation of den-
dendritic cell survival. Eur. J. Immunol. dritic cells leads to up-regulation
30:261219 of cellular FLICE-inhibitory protein
261. Anderson DM, Maraskovsky E, Bil- and concomitant down-regulation of
lingsley WL, Dougall WC, Tometsko death ligand-mediated apoptosis. Blood
ME, Roux ER, Teepe MC, DuBose RF, 96:262831
Cosman D, Galibert L. 1997. A homo- 268. McLellan AD, Terbeck G, Mengling
logue of the TNF receptor and its ligand T, Starling GC, Kiener PA, Gold R,
enhance T-cell growth and dendritic-cell Brocker EB, Leverkus M, Kampgen E.
function. Nature 390:17579 2000. Differential susceptibility to CD95
262. Wong BR, Josien R, Lee SY, Sauter B, (Apo-1/Fas) and MHC class II- induced
Li HL, Steinman RM, Choi Y. 1997. apoptosis during murine dendritic cell
TRANCE (tumor necrosis factor [TNF]- development. Cell Death Differ. 7:933
related activation-induced cytokine), a 38
new TNF family member predominantly 269. Bladergroen BA, Strik MC, Bovenschen
expressed in T cells, is a dendritic cell- N, van Berkum O, Scheffer GL, Meijer
specific survival factor. J. Exp. Med. 186: CJ, Hack CE, Kummer JA. 2001. The
207580 granzyme B inhibitor, protease inhibitor
263. Koch F, Heufler C, Kampgen E, 9, is mainly expressed by dendritic cells
Schneeweiss D, Bock G, Schuler G. and at immune-privileged sites. J. Im-
1990. Tumor necrosis factor alpha main- munol. 166:321825
tains the viability of murine epidermal 270. Medema JP, Schouurhuis D, Van Ton-
Langerhans cells in culture, but in con- geren J, De Jong J, Bres S, Toes R,
trast to granulocyte/macrophage colony- Toebes M, Schumacher T, Blader-
stimulating factor, without inducing Groen B, Ossendorp F, Kummer J, Me-
their functional maturation. J. Exp. Med. lief C, Offringa R. 2001. Expression of
171:15971 the serpin SPI-6 protects dendritic cells
264. Wang B, Fujisawa H, Zhuang L, Kondo from T lymphocyte-induced apoptosis:
S, Shivji GM, Kim CS, Mak TW, differential modulation by T-helper type
Sauder DN. 1997. Depressed Langer- 1 and type 2 cells. Presented at Keystone
hans cell migration and reduced con- Conference, Taos
tact hypersensitivity response in mice 271. Mackensen A, Drager R, Schlesier M,
lacking TNF receptor p75. J. Immunol. Mertelsmann R, Lindemann A. 2000.
159:614855 Presence of IgE antibodies to bovine
265. Funk JO, Walczak H, Voigtlander C, serum albumin in a patient developing
Berchtold S, Baumeister T, Rauch P, anaphylaxis after vaccination with hu-
Rossner S, Steinkasserer A, Schuler G, man peptide-pulsed dendritic cells. Can-
Lutz MB. 2000. Cutting edge: resistance cer Immunol. Immunother. 49:15256
to apoptosis and continuous prolifera- 272. Morse MA, Nair S, Fernandez-Casal M,
tion of dendritic cells deficient for TNF Deng Y, St Peter M, Williams R, Hob-
receptor-1. J. Immunol. 165:479296 eika A, Mosca P, Clay T, Cumming
266. Ashany D, Savir A, Bhardwaj N, Elkon RI, Fisher E, Clavien P, Proia AD,
KB. 1999. Dendritic cells are resistant Niedzwiecki D, Caron D, Lyerly HK.
2000. Preoperative mobilization of SC, Haines AM, Amir A, Husain R,

circulating dendritic cells by Flt3 lig- Doshi R, Young LS, Mountain A. 2000.
and administration to patients with Efficient nonviral transfection of den-
metastatic colon cancer. J. Clin. Oncol. dritic cells and their use for in vivo im-
18:388393 munization. Nat. Biotechnol. 18:1273
273. Choudhury BA, Liang JC, Thomas EK, 78
Flores-Romo L, Xie QS, Agusala K, 280. Gong J, Avigan D, Chen D, Wu Z,
Sutaria S, Sinha I, Champlin RE, Clax- Koido S, Kashiwaba M, Kufe D. 2000.
ton DF. 1999. Dendritic cells derived in Activation of antitumor cytotoxic T lym-
vitro from acute myelogenous leukemia phocytes by fusions of human dendritic
cells stimulate autologous, antileukemic cells and breast carcinoma cells. Proc.
T-cell responses. Blood 93:78086 Natl. Acad. Sci. USA 97:271518
274. Charbonnier A, Gaugler B, Sainty D, 281. Shu S, Cohen P. 2001. Tumor-dendritic

Lafage-Pochitaloff M, Olive D. 1999. cell fusion technology and immunother-
Human acute myeloblastic leukemia apy strategies. J. Immunother. 24:99
cells differentiate in vitro into mature 100

dendritic cells and induce the differen- 282. Celluzzi CM, Mayordomo JI, Storkus
tiation of cytotoxic T cells against au- WJ, Lotze MT, Falo LD Jr. 1996. Pep-
tologous leukemias. Eur. J. Immunol. tide-pulsed dendritic cells induce anti-
29:256778 gen-specific CTL-mediated protective
275. Eibl B, Ebner S, Duba C, Bock G, Ro- tumor immunity. J. Exp. Med. 183:283
mani N, Erdel M, Gachter A, Nieder- 87
wieser D, Schuler G. 1997. Dendritic 283. Porgador A, Snyder D, Gilboa E. 1996.
cells generated from blood precursors of Induction of antitumor immunity using
chronic myelogenous leukemia patients bone marrow-generated dendritic cells.
carry the Philadelphia translocation and J. Immunol. 156:291826
can induce a CML-specific primary cy- 284. Zitvogel L, Mayordomo JI, Tjandrawan
totoxic T-cell response. Genes Chromo- T, DeLeo AB, Clarke MR, Lotze MT,
somes Cancer 20:21523 Storkus WJ. 1996. Therapy of murine
276. Boczkowski D, Nair SK, Snyder D, tumors with tumor peptide-pulsed den-
Gilboa E. 1996. Dendritic cells pulsed dritic cells: dependence on T cells, B7
with RNA are potent antigen-presenting costimulation, and T helper cell 1-asso-
cells in vitro and in vivo. J. Exp. Med. ciated cytokines. J. Exp. Med. 183:8797
184:46572 285. Alters SE, Gadea JR, Philip R. 1997.
277. Nair SK, Heiser A, Boczkowski D, Ma- Immunotherapy of cancer. Generation of
jumdar A, Naoe M, Lebkowski JS, Vie- CEA specific CTL using CEA peptide
weg J, Gilboa E. 2000. Induction of pulsed dendritic cells. Adv. Exp. Med.
cytotoxic T cell responses and tumor im- Biol. 417:51924
munity against unrelated tumors using 286. Tjandrawan T, Martin DM, Maeurer
telomerase reverse transcriptase RNA MJ, Castelli C, Lotze MT, Storkus WJ.
transfected dendritic cells. Nat. Med. 1998. Autologous human dendriphages
6:101117 pulsed with synthetic or natural tumor
278. Jenne L, Schuler G, Steinkasserer A. peptides elicit tumor-specific CTLs in
2001. Viral vectors for dendritic cell- vitro. J. Immunother. 21:14957
based immunotherapy. Trends Immunol. 287. Bakker AB, Marland G, de Boer
22:1027 AJ, Huijbens RJ, Danen EH, Adema
279. Irvine AS, Trinder PK, Laughton DL, GJ, Figdor CG. 1995. Generation of
Ketteringham H, McDermott RH, Reid antimelanoma cytotoxic T lymphocytes
from healthy donors after presentation and protective immunity. Cell 92:535
of melanoma-associated antigen-derived 45
epitopes by dendritic cells in vitro. Can- 295. Blake N, Lee S, Redchenko I, Thomas
cer Res. 55:533034 W, Steven N, Leese A, Steigerwald-
288. Subklewe M, Chahroudi A, Bickham Mullen P, Kurilla MG, Frappier L,
K, Larsson M, Kurilla MG, Bhardwaj Rickinson A. 1997. Human CD8+ T cell
N, Steinman RM. 1999. Presentation responses to EBV EBNA1: HLA class I
of Epstein-Barr virus latency antigens to presentation of the (Gly-Ala)-containing
CD8(+), interferon-gamma-secreting, T protein requires exogenous processing.
lymphocytes. Eur. J. Immunol. 29:3995 Immunity 7:791802
4001 296. Dhodapkar MV, Steinman RM, Sapp
289. Amoscato AA, Prenovitz DA, Lotze M, Desai H, Fossella C, Krasovsky J,
MT. 1998. Rapid extracellular degra- Donahoe SM, Dunbar PR, Cerundolo
dation of synthetic class I peptides V, Nixon DF, Bhardwaj N. 1999. Rapid
by human dendritic cells. J. Immunol. generation of broad T-cell immunity in
161:402332 humans after a single injection of mature

290. Ludewig B, McCoy K, Pericin M, dendritic cells. J. Clin. Invest. 104:173
Ochsenbein AF, Dumrese T, Odermatt 80
B, Toes RE, Melief CJ, Hengartner 297. Zitvogel L, Angevin E, Tursz T. 2000.
H, Zinkernagel RM. 2001. Rapid pep- Dendritic cell-based immunotherapy of
tide turnover and inefficient presenta- cancer. Ann. Oncol. 11 (Suppl.) 3:199
tion of exogenous antigen critically limit 205
the activation of self-reactive CTL by 298. Nestle FO, Alijagic S, Gilliet M, Sun Y,
dendritic cells. J. Immunol. 166:3678 Grabbe S, Dummer R, Burg G, Scha-
87 dendorf D. 1998. Vaccination of mel-
291. Jager E, Ringhoffer M, Karbach J, anoma patients with peptide- or tumor
Arand M, Oesch F, Knuth A. 1996. lysate-pulsed dendritic cells. Nat. Med.
Inverse relationship of melanocyte dif- 4:32832
ferentiation antigen expression in 299. Murphy GP, Tjoa BA, Simmons SJ,
melanoma tissues and CD8+ cytotoxic- Ragde H, Rogers M, Elgamal A, Kenny
T-cell responses: evidence for immuno- GM, Troychak MJ, Salgaller ML,
selection of antigen-loss variants in Boynton AL. 1999. Phase II prostate
vivo. Int. J. Cancer 66:47076 cancer vaccine trial: report of a study
292. Riker A, Cormier J, Panelli M, Kam- involving 37 patients with disease re-
mula U, Wang E, Abati A, Fetsch P, Lee currence following primary treatment.
KH, Steinberg S, Rosenberg S, Marin- Prostate 39:5459
cola F. 1999. Immune selection af- 300. Panelli MC, Wunderlich J, Jeffries J,
ter antigen-specific immunotherapy of Wang E, Mixon A, Rosenberg SA,
melanoma. Surgery 126:11220 Marincola FM. 2000. Phase 1 study in
293. Albert ML, Darnell JC, Bender A, Fran- patients with metastatic melanoma of
cisco LM, Bhardwaj N, Darnell RB. immunization with dendritic cells pre-
1998. Tumor-specific killer cells in para- senting epitopes derived from the
neoplastic cerebellar degeneration. Nat. melanoma-associated antigens MART-1
Med. 4:132124 and gp100. J. Immunother. 23:48798
294. Shen H, Miller JF, Fan X, Kolwyck D, 301. Reichardt VL, Okada CY, Liso A, Ben-
Ahmed R, Harty JT. 1998. Compartmen- ike CJ, Stockerl-Goldstein KE, Engle-
talization of bacterial antigens: differen- man EG, Blume KG, Levy R. 1999.
tial effects on priming of CD8 T cells Idiotype vaccination using dendritic
cells after autologous peripheral blood a vaccine with peptide-pulsed dendritic

stem cell transplantation for multiple cells generated in vitro from CD34(+)
myelomaa feasibility study. Blood hematopoietic progenitor cells. Int. J.
93:241119 Cancer 86:38592
302. Small EJ, Fratesi P, Reese DM, Strang 307. Kugler A, Stuhler G, Walden P, Zoller
G, Laus R, Peshwa MV, Valone FH. G, Zobywalski A, Brossart P, Trefzer U,
2000. Immunotherapy of hormone- Ullrich S, Muller CA, Becker V, Gross
refractory prostate cancer with antigen- AJ, Hemmerlein B, Kanz L, Muller GA,
loaded dendritic cells. J. Clin. Oncol. 18: Ringert RH. 2000. Regression of human
3894903 metastatic renal cell carcinoma after vac-
303. Fong L, Hou Y, Rivas A, Benike C, cination with tumor cell-dendritic cell
Yuen A, Fisher GA, Davis MM, Engle- hybrids. Nat. Med. 6:33236
man EG. 2001. Altered peptide ligand 308. Kundu SK, Engleman E, Benike C,
vaccination with Flt3 ligand expanded Shapero MH, Dupuis M, van Schooten
dendritic cells for tumor immunother- WC, Eibl M, Merigan TC. 1998. A pi-
apy. Proc. Natl. Acad. Sci. USA 98:8809 lot clinical trial of HIV antigen-pulsed
14 allogeneic and autologous dendritic cell
304. Thurner B, Haendle I, Roder C, Dieck- therapy in HIV-infected patients. AIDS
mann D, Keikavoussi P, Jonuleit H, Res. Hum. Retroviruses 14:55160
Bender A, Maczek C, Schreiner D, von 309. Zitvogel L, Regnault A, Lozier A, Wol-
den Driesch P, Brocker EB, Steinman fers J, Flament C, Tenza D, Ricciardi-
RM, Enk A, Kampgen E, Schuler G. Castagnoli P, Raposo G, Amigorena S.
1999. Vaccination with mage-3A1 pep- 1998. Eradication of established murine
tide-pulsed mature, monocyte-derived tumors using a novel cell-free vaccine:
dendritic cells expands specific cyto- dendritic cell-derived exosomes. Nat.
toxic T cells and induces regression of Med. 4:594600
some metastases in advanced stage IV 310. Thery C, Regnault A, Garin J, Wolfers J,
melanoma. J. Exp. Med. 190:166978 Zitvogel L, Ricciardi-Castagnoli P, Ra-
305. Holtl L, Rieser C, Papesh C, Ramoner poso G, Amigorena S. 1999. Molecular
R, Herold M, Klocker H, Radmayr C, characterization of dendritic cell-derived
Stenzl A, Bartsch G, Thurnher M. 1999. exosomes. Selective accumulation of the
Cellular and humoral immune responses heat shock protein hsc73. J. Cell Biol.
in patients with metastatic renal cell car- 147:599610
cinoma after vaccination with antigen- 311. Thery C, Boussac M, Veron P, Ric-
pulsed dendritic cells. J. Urol. 161:777 ciardi-Castagnoli P, Raposo G, Garin J,
82 Amigorena S. 2001. Proteomic analysis
306. Mackensen A, Herbst B, Chen JL, Koh- of dendritic cell-derived exosomes: a se-
ler G, Noppen C, Herr W, Spagnoli creted subcellular compartment distinct
GC, Cerundolo V, Lindemann A. 2000. from apoptotic vesicles. J. Immunol.
Phase I study in melanoma patients of 166:730918
P1: FRK
February 1, 2002 12:47 Annual Reviews AR152-FM
Annual Review of Immunology

Volume 20, 2002
CONTENTS
FRONTISPIECE, Charles A. Janeway, Jr. xiv
A TRIP THROUGH MY LIFE WITH AN IMMUNOLOGICAL THEME,
Charles A. Janeway, Jr. 1

THE B7 FAMILY OF LIGANDS AND ITS RECEPTORS: NEW PATHWAYS
FOR COSTIMULATION AND INHIBITION OF IMMUNE RESPONSES,
Beatriz M. Carreno and Mary Collins 29

MAP KINASES IN THE IMMUNE RESPONSE, Chen Dong, Roger J. Davis,
and Richard A. Flavell 55
PROSPECTS FOR VACCINE PROTECTION AGAINST HIV-1 INFECTION
AND AIDS, Norman L. Letvin, Dan H. Barouch, and David C. Montefiori 73
T CELL RESPONSE IN EXPERIMENTAL AUTOIMMUNE
ENCEPHALOMYELITIS (EAE): ROLE OF SELF AND CROSS-REACTIVE
ANTIGENS IN SHAPING, TUNING, AND REGULATING THE
AUTOPATHOGENIC T CELL REPERTOIRE, Vijay K. Kuchroo,
Ana C. Anderson, Hanspeter Waldner, Markus Munder, Estelle Bettelli,
and Lindsay B. Nicholson 101
NEUROENDOCRINE REGULATION OF IMMUNITY, Jeanette I. Webster,
Leonardo Tonelli, and Esther M. Sternberg 125
MOLECULAR MECHANISM OF CLASS SWITCH RECOMBINATION:
LINKAGE WITH SOMATIC HYPERMUTATION, Tasuku Honjo, Kazuo
Kinoshita, and Masamichi Muramatsu 165
INNATE IMMUNE RECOGNITION, Charles A. Janeway, Jr. and Ruslan
Medzhitov 197
KIR: DIVERSE, RAPIDLY EVOLVING RECEPTORS OF INNATE AND
ADAPTIVE IMMUNITY, Carlos Vilches and Peter Parham 217
ORIGINS AND FUNCTIONS OF B-1 CELLS WITH NOTES ON THE ROLE
OF CD5, Robert Berland and Henry H. Wortis 253
E PROTEIN FUNCTION IN LYMPHOCYTE DEVELOPMENT, Melanie
W. Quong, William J. Romanow, and Cornelis Murre 301
LYMPHOCYTE-MEDIATED CYTOTOXICITY, John H. Russell and
Timothy J. Ley 323
SIGNAL TRANSDUCTION MEDIATED BY THE T CELL ANTIGEN
x
P1: FRK
February 1, 2002 12:47 Annual Reviews AR152-FM
CONTENTS xi
RECEPTOR: THE ROLE OF ADAPTER PROTEINS, Lawrence E. Samelson 371

INTERACTION OF HEAT SHOCK PROTEINS WITH PEPTIDES AND
ANTIGEN PRESENTING CELLS: CHAPERONING OF THE INNATE AND
ADAPTIVE IMMUNE RESPONSES, Pramod Srivastava 395
CHROMATIN STRUCTURE AND GENE REGULATION IN THE IMMUNE
SYSTEM, Stephen T. Smale and Amanda G. Fisher 427
PRODUCING NATURES GENE-CHIPS: THE GENERATION OF PEPTIDES
FOR DISPLAY BY MHC CLASS I MOLECULES, Nilabh Shastri, Susan
Schwab, and Thomas Serwold 463
THE IMMUNOLOGY OF MUCOSAL MODELS OF INFLAMMATION, Warren
Strober, Ivan J. Fuss, and Richard S. Blumberg 495

T CELL MEMORY, Jonathan Sprent and Charles D. Surh 551
GENETIC DISSECTION OF IMMUNITY TO MYCOBACTERIA: THE HUMAN

MODEL, Jean-Laurent Casanova and Laurent Abel 581
ANTIGEN PRESENTATION AND T CELL STIMULATION BY DENDRITIC
CELLS, Pierre Guermonprez, Jenny Valladeau, Laurence Zitvogel, Clotilde
Thery, and Sebastian Amigorena 621
NEGATIVE REGULATION OF IMMUNORECEPTOR SIGNALING, Andre
Veillette, Sylvain Latour, and Dominique Davidson 669
CPG MOTIFS IN BACTERIAL DNA AND THEIR IMMUNE EFFECTS,
Arthur M. Krieg 709
PROTEIN KINASE C IN T CELL ACTIVATION, Noah Isakov and Amnon
Altman 761
RANK-L AND RANK: T CELLS, BONE LOSS, AND MAMMALIAN
EVOLUTION, Lars E. Theill, William J. Boyle, and Josef M. Penninger 795
PHAGOCYTOSIS OF MICROBES: COMPLEXITY IN ACTION, David
M. Underhill and Adrian Ozinsky 825
STRUCTURE AND FUNCTION OF NATURAL KILLER CELL RECEPTORS:
MULTIPLE MOLECULAR SOLUTIONS TO SELF, NONSELF
DISCRIMINATION, Kannan Natarajan, Nazzareno Dimasi, Jian Wang,
Roy A. Mariuzza, and David H. Margulies 853
INDEXES
Subject Index 887
Cumulative Index of Contributing Authors, Volumes 120 915
Cumulative Index of Chapter Titles, Volumes 120 925
ERRATA
An online log of corrections to Annual Review of Immunology
chapters (if any, 1997 to the present) may be found
at http://immunol.annualreviews.org/

Antigen Presentation and T Cell Stimulation by Dendritic Cells

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Antigen Presentation and T Cell Stimulation by Dendritic Cells

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Annu. Rev. Immunol. 2002. 20:62167

ANTIGEN PRESENTATION AND T CELL

Institut Curie; e-mail: Pierre.Guermonprez@curie.fr, Jenny.Valladeau@curie.fr,

Key Words T cell priming, immunotherapy, MHC, endocytosis, class presentation

Dendritic cells represent a heterogenous cell population, residing in most periph-

622 GUERMONPREZ ET AL.

specific dendritic cell surface receptors. T lymphocytes, through CD40-dependent

in the expression of chemokine receptors and adhesion molecules, as well as pro-

ANTIGEN AND PATHOGEN RECOGNITION

ANTIGEN PRESENTATION BY DENDRITIC CELLS 623

Receptor-mediated endocytosis allows the uptake of macromolecules through spe-

ognized by a family of adaptors responsible for the recruitment of clathrin lattices

FC, COMPLEMENT, HEAT SHOCK PROTEINS, AND SCAVENGER RECEPTORS Mouse

624 GUERMONPREZ ET AL.

C-TYPE LECTIN FAMILY C-type lectins bind ligands in a Ca++-dependent manner.

Phagocytosis and Macropinocytosis

ANTIGEN PRESENTATION BY DENDRITIC CELLS 625

PHAGOCYTOSIS OF PATHOGENS Immature dendritic cells were reported to phago-

PHAGOCYTOSIS OF APOPTOTIC AND NECROTIC BODIES Immature dendritic cells

infection (influenza, vaccinia, measles, EBV, HIV1), X-ray-, - or UV-irradiation,

Regulation of Endocytosis in Dendritic Cells

626 GUERMONPREZ ET AL.

(e.g., MMR/FcR) and the downmodulation of both macropinocytosis and phago-

Pathogens Use Dendritic Cell Endocytic Receptors

INDUCTION OF DENDRITIC CELL MATURATION

ANTIGEN PRESENTATION BY DENDRITIC CELLS 627

triggers effector cells of inflammation, including macrophages and polymorphonu-

In this chapter, we briefly describe the receptors involved in the induction of

Receptors for Dendritic Cell Activation

628 GUERMONPREZ ET AL.

Signal Transduction and Dendritic Cell Maturation

ANTIGEN PRESENTATION BY DENDRITIC CELLS 629

of immature dendritic cells induces maturation through the activation of NF-B

630 GUERMONPREZ ET AL.

ANTIGEN PRESENTATION AND CROSS-PRESENTATION

mentalization of MHC class I and II biogenesis results in the loading of exogenous

MHC Class IRestricted Antigen Presentation

ANTIGEN PRESENTATION BY DENDRITIC CELLS 631

the immunoproteasome, which becomes the main type of proteasome in mature

WHICH INTERNALIZATION PATHWAYS LEAD TO CROSS PRESENTATION? Exogenous

MHC class I molecules. Macropinocytosis allows receptor-independent cross pre-

antigens to latex beads, thus forcing internalization by phagocytosis, strongly in-

632 GUERMONPREZ ET AL.

HOW ARE INTERNALIZED ANTIGENS CROSS-PRESENTED? Two main intracellular

ANTIGEN PRESENTATION BY DENDRITIC CELLS 633

or -independent pathways according to the sequence context of the peptides (16).

MHC Class IIRestricted Antigen Presentation

634 GUERMONPREZ ET AL.

First, antigen degradation is inefficient in immature dendritic cells, and inter-

ANTIGEN PRESENTATION BY DENDRITIC CELLS 635

CD1-Restricted Antigen Presentation

636 GUERMONPREZ ET AL.

counteract the CD1d-dependent NKT cell stimulation (176).

T CELL STIMULATION BY DENDRITIC CELLS

Priming and Tolerization

ANTIGEN PRESENTATION BY DENDRITIC CELLS 637

was sufficient to trigger V-specific deletion of T cells by a retrovirally encoded

Cross Priming and Cross Tolerization

tation and Cross Presentation) (102). Examples of productive immunization against

638 GUERMONPREZ ET AL.

Dendritic Cell Subsets and T Cell Priming