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J Virol, May 1998, p. 3812-3818, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Dendritic Cells Efficiently Induce Protective
Antiviral Immunity
Burkhard
Ludewig,*
Stephan
Ehl,
Urs
Karrer,
Bernhard
Odermatt,
Hans
Hengartner, and
Rolf M.
Zinkernagel
Institute of Experimental Immunology, CH-8091
Zürich, Switzerland
Received 8 October 1997/Accepted 12 January 1998
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ABSTRACT |
Cytotoxic T lymphocytes (CTL) are essential for effective immunity
to various viral infections. Because of the high speed of viral
replication, control of viral infections imposes demanding functional
and qualitative requirements on protective T-cell responses. Dendritic
cells (DC) have been shown to efficiently acquire, transport, and
present antigens to naive CTL in vitro and in vivo. In this study, we
assessed the potential of DC, either pulsed with the lymphocytic
choriomeningitis virus (LCMV)-specific peptide GP33-41 or
constitutively expressing the respective epitope, to induce LCMV-specific antiviral immunity in vivo. Comparing different application routes, we found that only 100 to 1,000 DC had to reach the
spleen to achieve protective levels of CTL activation. The DC-induced
antiviral immune response developed rapidly and was long lasting.
Already at day 2 after a single intravenous immunization with high
doses of DC (1 × 105 to 5 × 105),
mice were fully protected against LCMV challenge infection, and direct
ex vivo cytotoxicity was detectable at day 4 after DC immunization. At
day 60, mice were still protected against LCMV challenge infection.
Importantly, priming with DC also conferred protection against
infections in which the homing of CTL into peripheral organs is
essential: DC-immunized mice rapidly cleared an infection with
recombinant vaccinia virus-LCMV from the ovaries and eliminated LCMV
from the brain, thereby avoiding lethal choriomeningitis. A comparison
of DC constitutively expressing the GP33-41 epitope with exogenously
peptide-pulsed DC showed that in vivo CTL priming with peptide-loaded
DC is not limited by turnover of peptide-major histocompatibility
complex class I complexes. We conclude that the priming of antiviral
CTL responses with DC is highly efficient, rapid, and long lasting.
Therefore, the use of DC should be considered as an efficient means of
immunization for antiviral vaccination strategies.
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INTRODUCTION |
Dendritic cells (DC) derived from
bone marrow (bmDC) are found as antigen-presenting cells (APC)
scattered throughout nonlymphoid tissues. After antigen acquisition and
processing, DC migrate via lymph vessels or blood to the T-cell areas
of regional secondary lymphoid organs, where they present major
histocompatibility complex (MHC) class I- and II-restricted peptides to
naive T cells (42). The combination of these properties,
i.e., antigen uptake, processing, transport, and presentation, makes DC
particularly suitable as vehicles for antigens in immunotherapy. DC
pulsed with tumor-derived peptides (29, 39, 46),
tumor-associated proteins (16), or tumor cell RNA
(9) have been shown to activate tumor-specific cytotoxic T
lymphocytes (CTL) and to reduce tumor load.
The high efficiency of DC in eliciting T-cell responses against
infectious agents has been demonstrated with different experimental systems; e.g., DC can prime immune responses against mycobacteria (19), Borrelia burgdorferi (30), and
Leishmania major (31). Induction of
virus-specific CTL by DC in vitro with virus peptides or virus proteins
has been shown for influenza virus (27), Sendai virus
(22), and human immunodeficiency virus (26). In
vivo, adoptive transfer of human immunodeficiency virus peptide-pulsed DC (43) or hepatitis B virus protein-pulsed DC
(10) elicits virus-specific CTL responses. Whether these CTL
responses reflect the ability to provide antiviral protection in vivo,
however, has not been addressed in these studies. This aspect is
particularly important in viral infections, since the high speed of
replication and amplification imposes demanding quantitative and
functional requirements on protective CTL responses (6, 15,
24).
Infection of mice with lymphocytic choriomeningitis virus (LCMV) is a
well-characterized model system for studying CTL responses in vivo.
Control of an acute virus infection is almost exclusively achieved by
CD8+ T-cell-mediated, perforin-dependent cytotoxicity
(21). Various in vivo and in vitro assays with graded
sensitivities allow assessment of the efficiency and biological
relevance of an immunization strategy (5, 12). In addition,
several CTL immunization strategies, including peptide vaccination
(1, 2, 41) and priming with recombinant vaccinia virus
(17), have been well characterized for this experimental
system. Therefore, the study of DC-induced immunity against LCMV
infection allows an interesting qualitative comparison with other
vaccination approaches.
In the present study, we evaluated the potential of DC to induce
CTL-mediated antiviral immunity in vivo using bone marrow-derived DC
either pulsed with peptides or obtained from transgenic mice constitutively expressing the immunodominant epitope (GP33-41; hereafter referred to as GP33) of the LCMV (WE strain) glycoprotein (14a). Immunization with these cells allowed us to address
the following questions. (i) How many DC are needed to induce
protection against a viral infection? (ii) What are the in vivo
kinetics of DC-induced CTL activation? (iii) Do DC-induced antiviral
CTL emigrate to peripheral tissues to resolve viral infections, and are
they protective against virus-induced immunopathology? (iv) Does
turnover of peptide-MHC class I complexes influence DC immunogenicity?
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MATERIALS AND METHODS |
Mice.
C57BL/6 mice were obtained from the Institut für
Labortierkunde (University of Zürich, Zürich, Switzerland).
Transgenic mice expressing the LCMV GP33 epitope ubiquitously in all
tissues were generated by use of a construct containing the
H-2Kb regulatory elements and the coding
sequence for amino acids 1 to 60 of the LCMV glycoprotein (H8 mice)
(14a). The transgene construct was injected into fertilized
C57BL/6 oocytes. All animals were kept under specific-pathogen-free
conditions.
Viruses and cell lines.
LCMV strain WE was originally
obtained from F. Lehmann-Grube (Hamburg, Germany) and propagated on
L929 cells. Recombinant vaccinia virus expressing LCMV glycoprotein
(Vacc-G2) was obtained from D. H. Bishop (Oxford, United Kingdom)
and grown on BSC40 cells. Viruses were titrated as described previously
with MC-57 cells for LCMV (8) and BSC40 cells for Vacc-G2
(17) cells. EL-4 (H-2b), a thymoma
cell line, was used as the target cell line.
Antibodies and peptides.
Supernatants from the following
monoclonal antibody-producing hybridomas (American Type Culture
Collection) were used: rat anti-mouse CD4 (YTS191.1), rat anti-mouse
CD8 (YTS169.4.2), rat anti-mouse CD45R (RA3-3A1/6.1), and rat
anti-mouse I-Abd (B21-2).
LCMV GP33 (KAVYNFATM) (37) was synthesized by the
solid-phase method and purchased from Neosystem Laboratoire
(Strasbourg, France). To prevent dimer formation, the original cysteine
at anchor position 41 in LCMV GP33 was replaced by methionine.
Preparation of DC.
For generation of DC from C57BL/6 and H8
mouse bone marrow cultures (H8-bmDC and B6-bmDC, respectively), the
procedure of Inaba et al. (20) was used, with minor
modifications. Briefly, bone marrow was flushed from femurs and tibias
and subsequently depleted of erythrocytes with ammonium chloride. Bone
marrow cells were depleted of lymphocytes, B cells, and
I-Abd+ cells by use of a cocktail of monoclonal
antibodies (YTS191.1, YTS169.4.2, RA3-3A1/6.1, and B21-2) and goat
anti-rat immunoglobulin G-coated Dynabeads (Dynal, Oslo, Norway). Cells
were plated at 0.5 × 106/ml in RPMI 1640 supplemented
with 5% fetal calf serum (FCS), penicillin-streptomycin, 10 ng of
recombinant murine granulocyte-macrophage colony-stimulating factor
(kindly supplied by Sandoz, Vienna, Austria) per ml, and interleukin
4-containing supernatant from cell line X63-IL4 (kindly provided by M. Kopf, Basel, Switzerland) at a final concentration of 100 ng/ml. At
days 2 and 4 of culturing, 50% of the supernatant was removed and
replenished with fresh medium, and fresh cytokines were added. At day
6, nonadherent cells were collected and further purified over
metrizamide (14.5% in RPMI 1640 containing 5% FCS) (Sigma) to remove
cell debris and high-density cells.
The resulting bmDC populations showed a high purity, with 80 to 90% of
the cells showing the distinct stellate DC morphology.
More than 85%
of the cells were CD11b
+ and showed strong expression of
MHC class I and II antigens (5
to 10 times higher than that of naive B
cells). A total of 70
to 85% of the cells were positive for CD80,
CD86, and the DC marker
NLDC-145, as determined by flow cytometry (data
not shown). The
differentiation stage of these DC was immature, as
judged by the
ability to take up, process, and present native viral
antigens
to transgenic T-helper cells (data not shown). Furthermore,
the
activation of bmDC with tumor necrosis factor induced significant
upregulation of MHC and costimulatory antigens (data not shown).
To enrich for splenic DC (sDC), spleens were digested with collagenase
and ground through a stainless steel screen with a
sterile syringe
plunger. Cells were resuspended in RPMI 1640 supplemented
with 10% FCS
and antibiotics at 2 × 10
7/ml and cultured on 10-cm
tissue culture dishes for 90 min. Nonadherent
cells were removed by
washing with phosphate-buffered saline,
and adherent cells were
cultured overnight in RPMI 1640 supplemented
with 10% FCS, 10 ng of
granulocyte-macrophage colony-stimulating
factor per ml, and 100 ng of
interleukin 4 per ml. Nonadherent,
low-density cells were separated
with metrizamide. Cell suspensions
prepared in this way contained 30 to
60% DC, as determined by
morphology and flow cytometry.
Peptide treatment and immunization of mice.
Purified C57BL/6
bmDC and sDC (1 × 106 to 5 × 106)
were resuspended in 0.5 ml of medium containing GP33 at a concentration
of 10
6 M and incubated for 60 min at 37°C on a rocking
platform. Cells were washed three times with balanced salt solution
(BSS) and resuspended at 2 × 106/ml in BSS, and
serial 10-fold dilutions were made. DC were injected in a volume of 0.5 ml intravenously (i.v.), 0.05 ml subcutaneously (s.c.) at the base of
the tail, or 0.02 ml directly into the spleen.
Cytotoxicity assays.
Spleen cells (4 × 106/well) from primed mice were restimulated for 5 days in
24-well tissue culture plates with 2 × 106
GP33-labeled, irradiated (3,000 rads) spleen cells or with 2 × 105 LCMV-infected, irradiated peritoneal macrophages in
Iscove's modification of Dulbecco's medium (IMDM) supplemented with
10% FCS, penicillin- streptomycin, and 0.001 M 2-mercaptoethanol. Restimulated spleen effector cells from one well were resuspended in 1 ml of minimal essential medium containing 2% FCS, and serial threefold
dilutions were made (indicated in figures as dilution of culture). For
detection of primary ex vivo cytotoxicity, effector cell suspensions
were prepared from spleens of immunized mice at various times after
priming. EL-4 cells were pulsed with LCMV GP33 (106 M,
1.5 h, 37°C) and used in a standard 5-h 51Cr release
assay or in an overnight (15-h) assay. 51Cr-labeled
nonpulsed EL-4 cells served as controls. Spontaneous release was always
below 19% for 5-h assays and below 29% for overnight assays.
CTLp assay.
Quantification of GP33-specific precursor CTL
(CTLp) per spleen was performed by limiting-dilution analysis as
described previously (32). Responder spleen cells were
titrated and cultured with 104 LCMV-infected, irradiated
peritoneal macrophages and 105 irradiated feeder spleen
cells in IMDM-10% FCS and 10% concanavalin A supernatant in 16 wells
per dilution step. After 6 days of culturing, cytotoxicity was tested
on GP33 containing loaded or unloaded EL-4 cells in a 51Cr
release assay, and CTLp frequencies were calculated (44).
CFSE labeling of DC and immunohistochemistry.
The
fluorescent dye CFSE (5- and 6-carboxyfluorescein diacetate
succinimidyl ester) was purchased from Molecular Probes (Eugene, Oreg.). DC were washed with BSS, resuspended at 106/ml in
BSS containing 0.5 µM CFSE, and incubated for 10 min at 37°C. After
the cells were labeled, FCS was added to a final concentration of 5%,
and the cells were washed twice. DC viability after CFSE labeling was
>95%, as determined by trypan blue exclusion. CFSE-labeled DC (3 × 106) were adoptively transferred into recipient mice.
Organs were removed at days 1 and 2, immersed in Hanks BSS, and snap
frozen in liquid nitrogen. Histological procedures were performed as described previously (33) with rabbit antifluorescein
antibody (Dako).
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RESULTS |
Migration pattern for bmDC.
Apart from their high stimulatory
capacity, the ability of DC to migrate to secondary lymphoid organs and
to home to T-cell areas is a prerequisite for efficient CTL priming. To
assess the homing pattern for bmDC, cells were labeled with the
fluorescent dye CFSE and adoptively transferred into C57BL/6 recipient
mice. One to 2 days later, several organs were analyzed for the
presence of the donor DC by immunohistochemistry. After i.v. injection, the majority of the CFSE-labeled bmDC homed to the T-cell areas of the
spleen (Fig. 1A). In the T-cell areas, DC
displayed the typical stellate morphology, thereby facilitating
interaction with a large number of T cells (Fig. 1B). Homing of bmDC to
paracortical areas of regional hepatic lymph nodes was also observed,
as previously described for rat blood DC (23). Transferred
DC were hardly detectable in other lymph nodes (mediastinal, inguinal,
and axillary) or other tissues (liver, lung, and kidney) (data not
shown). Thus, after i.v. injection, bmDC preferentially home to the
T-cell areas of the spleen.
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FIG. 1.
Homing of bmDC to splenic T-cell areas. bmDC (3 × 106) were CFSE labeled, injected i.v., and visualized in
cryostat sections with an antifluorescein antibody. (A) bmDC were found
in the interfollicular areas (follicles are marked with asterisks).
Magnification, ×40. (B) High magnification of boxed area in panel A
showing dense follicular area (asterisk), DC with typical stellate
morphology (arrow), and central artery (arrowhead). Magnification,
×350.
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How many DC are needed to induce antiviral immunity?
To
evaluate the antiviral priming capacity of DC, we used transgenic mice
expressing the first 60 amino acids of the LCMV glycoprotein under the
control of the H-2Kb promoter (H8 mice). This
characteristic results in the ubiquitous expression of the
immunodominant LCMV epitope GP33 (14a). When H8-bmDC were
used, external loading of DC with either peptide or protein was not
needed.
To determine the minimal number of DC required for the induction of
antiviral immunity and to assess the influence of DC migration
on the
efficiency of CTL induction, C57BL/6 mice (
H-2b)
were immunized with different doses of H8-bmDC by direct injection
into
the spleen (Fig.
2A), i.v. (Fig.
2B), or
s.c. (Fig.
2C).
Eight days later, spleens were harvested, and
cytotoxicity was
determined after restimulation for 5 days with
GP33-pulsed, irradiated
spleen cells. The minimal dose needed to elicit
a measurable CTL
response after intrasplenic injection was
approximately 100 H8-bmDC
(Fig.
2A); injection of lower numbers of DC
did not induce a CTL
response (data not shown). The minimal dose needed
to induce a
CTL response in the spleen after i.v. injection was
10
3 DC (Fig.
2B). In contrast, approximately 10 times more
H8-bmDC
had to be injected s.c. to reach similar levels of CTL
induction
in the spleen (Fig.
2C).
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FIG. 2.
Efficient priming of antiviral CTL by H8-bmDC. C57BL/6
mice were immunized by intrasplenic injection (A), i.v. (B), or s.c.
(C) with graded doses of H8-bmDC. Eight days later, induction of
GP33-specific CTL was tested. Spleen cells were restimulated in vitro
for 5 days with peptide-labeled, irradiated spleen cells. Specific
lysis was measured on GP33-labeled EL-4 target cells (closed symbols)
or EL-4 cells without peptide (open symbols). Spontaneous release was
<15%. Values for control mice immunized i.v. with 105
unlabeled B6-bmDC were 10, 7, 4, and 2% with labeled EL-4 cells and
12, 9, 4, and 1% with unlabeled EL-4 cells at the indicated culture
dilutions (from left to right), respectively.
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To test the protective capacity of H8-bmDC, mice immunized via
different routes with graded doses of H8-bmDC were challenged
with 200 PFU of LCMV strain WE i.v. at day 8 after DC priming.
Infection of
naive C57BL/6 mice caused high virus titers (>10
6
PFU/spleen), whereas mice immunized by intrasplenic injection
with
10
3 H8-bmDC were able to rapidly clear the virus (Fig.
3). Mice immunized
intrasplenically with
10
2 H8-bmDC showed a reduction of virus titers of about 2 to 4 log
units but did not completely clear the virus by day 4. Immunization
with 10
4 H8-bmDC i.v. or 10
5
H8-bmDC s.c. led to resistance against a low-dose LCMV challenge
infection, whereas injection of 10
3 H8-bmDC i.v. or
10
4 H8-bmDC s.c. only partially protected against a
low-dose challenge
infection (Fig.
3).
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FIG. 3.
Protection against low-dose LCMV challenge after priming
with H8-bmDC. Mice were immunized intrasplenically (i.spl.), i.v. or
s.c. (at the base of the tail) with the indicated number of H8-bmDC. At
day 8 postimmunization, protection against LCMV strain WE was tested.
Virus titers in spleens were determined 4 days after an i.v. challenge
with 200 PFU of LCMV strain WE. The detection limit is represented by
the broken line.
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The results described above suggested that low numbers of DC are
sufficient to induce an in vivo protective antiviral immune
response.
To assess how this response is reflected in an increase
in the number
of virus-specific CTLp in the spleen, mice were
injected with different
doses of H8-bmDC i.v. and a limiting-dilution
assay was performed 8 days later. With doses of 10
4 to 10
6 DC,
GP33-specific CTLp frequencies were always about 1:28,000
to 1:100,000
(Table
1). In LCMV memory mice (i.e.,
mice infected
with 10
2 PFU of LCMV WE at least 50 days
ago), CTLp frequencies were also
about 1:10,000 (Table
1)
(
3). Thus, in the LCMV system, DC
efficiently prime CTL to
levels of protective memory responses.
Rapid and long-lasting activation of CTL by DC.
To evaluate
the kinetics of CTL induction by DC, LCMV GP33-specific cytotoxicity
was monitored at different times after adoptive transfer of H8-bmDC
into C57BL/6 mice. DC-induced primary ex vivo cytotoxicity was weak, as
determined by a standard 5-h 51Cr release assay (Fig.
4). When the sensitivity of the assay was increased by prolongation of the incubation time to 15 h, direct ex vivo cytotoxicity became apparent 4 days after priming with a high
dose of DC (5 × 105) (Fig. 4A to D). Immunization
with a low dose of DC (104) led to detectable ex vivo
cytotoxicity beginning at day 6 (Fig. 4E to G).
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FIG. 4.
Kinetics of CTL induction after DC priming. Mice were
primed with either 5 × 105 (A to D) or
104 (E to G) H8-bmDC i.v. At different times after
immunization, ex vivo CTL activity of splenocytes was tested in a
51Cr release assay after 5 h (open symbols) or after
15 h (closed symbols) on GP33-labeled EL-4 target cells at the
indicated effector/target cell (E:T) ratios. Spontaneous release after
5 h was <12%; after 15 h, it was <29%. Nonspecific lysis
of unlabeled EL-4 target cells was always <5%. Values (percentages)
for an LCMV-infected control mouse (day 8) were 87, 74, 61, and 35%
for 5 h and 99, 98, 81, and 72% for 15 h at E:T ratios of
90, 30, 10, and 3, respectively.
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Challenging H8-bmDC-primed mice at different times after adoptive
transfer showed the rapid development and long duration
of the
antiviral immune response. Immunization with 10
5 H8-bmDC
completely protected mice against challenge with LCMV
infection as
early as 2 days after priming (Fig.
5).
With 10
4 H8-bmDC, protective immunity had not developed at
day 2 but was
detectable by day 8 postimmunization (Fig.
5). After 60 days,
mice immunized with 10
5 H8-bmDC were still fully
protected against LCMV challenge, whereas
immunization with
10
4 H8-bmDC was only partially protective. In addition,
CTLp frequencies
at day 60 after DC immunization were significantly
elevated (Table
1). Taken together, these results show that high
numbers of DC
rapidly induce a long-lasting protective immune response.
With
low numbers of DC, mounting of a fully protective immune response
was slightly delayed and protection waned faster.
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FIG. 5.
Rapid and long-lasting induction of protective immunity
by high doses of DC. Mice were immunized i.v. with different doses of
H8-bmDC and challenged with 200 PFU of LCMV strain WE i.v. after 2, 8, or 60 days. Virus titers in spleens were determined 4 days after
challenge infection. The broken line represents the detection limit.
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DC efficiently prime against peripheral virus challenge.
In
the experiments described above, antiviral CTL activity was determined
mainly in the spleen. Control of virus replication outside lymphoid
tissues (such as ovaries or brain) imposes much more stringent
requirements on virus-specific CTL (6, 24). To assess
whether DC-primed CTL exert their antiviral activity after homing to
peripheral organs in vivo, virus control in the ovaries after
intraperitoneal infection with LCMV glycoprotein-expressing vaccinia
virus (Vacc-G2) and in the brains after intracerebral challenge with
LCMV strain WE was studied. Immunization with 103 H8-bmDC
only partially protected against Vacc-G2 challenge, whereas higher
doses completely inhibited Vacc-G2 growth (Table
2). After intracerebral injection of LCMV
strain WE, naive mice started to show symptoms of immunopathological
choriomeningitis on day 7. Mice immunized with 103 H8-bmDC
and three of five mice immunized with 104 DC had already
developed disease by days 5 to 6, indicative of CTL preactivation
leading to enhanced immunopathology (34). Mice immunized
with 105 H8-bmDC and one of five mice immunized with
104 DC survived the intracerebral challenge with LCMV
strain WE without developing symptoms of choriomeningitis (Table 2).
Thus, DC-primed CTL rapidly eliminate virus from peripheral tissues,
thereby preventing later virus-induced, CTL-mediated immunopathology.
In vivo CTL priming with peptide-loaded DC is not influenced by
turnover of peptide-MHC class I complexes.
H8-bmDC constitutively
express the transgenic glycoprotein fragment and therefore the GP33
epitope. Thus, comparison of H8-bmDC with B6-bmDC and sDC from C57BL/6
mice (B6-sDC) loaded exogenously with GP33 peptide allowed us to assess
whether peptide turnover and decay influence the in vivo priming
capacities of DC. In preliminary experiments, peptide loading of
B6-bmDC was tested with activation of GP33-specific T-cell
receptor-transgenic T cells (36). Labeling of B6-bmDC with
GP33 for 1 h at a concentration of 106 M led to an
in vitro stimulatory capacity comparable to that of H8-bmDC (data not
shown). In vivo, immunization with GP33-labeled B6-bmDC or B6-sDC led
to induction of CTL responses (Fig. 6A) and protection against LCMV strain WE challenge (Fig. 6B) with the same
efficiency as H8-bmDC immunization.
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FIG. 6.
Priming efficiencies of H8-bmDC and of peptide-labeled
B6-sDC or B6-bmDC. (A) Mice were injected i.v. with 104
H8-bmDC or 104 GP33-labeled B6-sDC or B6-bmDC, and specific
CTL induction was determined as described in the legend to Fig. 2. (B)
Mice were immunized i.v. with different doses of H8-bmDC or
GP33-labeled B6-sDC or B6-bmDC and challenged with 200 PFU of LCMV
strain WE i.v. at day 8. Virus titers in spleens were determined 4 days
later.
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DISCUSSION |
In the present study, we evaluated the ability of DC to
induce CTL-mediated antiviral immunity in vivo. When administered i.v.,
as few as 1,000 to 10,000 DC, either constitutively expressing an MHC
class I-restricted epitope or exogenously loaded with peptide, led to
the induction of virus-specific CTL, protected against challenge LCMV
infection, and prevented viral replication in peripheral tissues.
The high efficiency of DC in priming T-cell responses against tumors
(16, 29, 39, 46) and nonviral infections (19, 30,
31) in vivo has been shown. CTL are crucial for effective immune
responses against various viral infections. Induction of antiviral CTL
responses by DC has been studied in vivo mainly with secondary in vitro
restimulation (10, 43). In the present study, we assessed
the induction of protective antiviral CTL responses by DC in vivo. The
infectious LCMV mouse model is particularly useful for this purpose, as
DC are infected during primary LCMV infection (11) and
well-established in vivo assays (5) allow investigation of
the extent of CTL priming. Furthermore, the finely graded sensitivity
of these assays facilitates comparison with other anti-LCMV
immunization strategies. Priming with peptide GP33 in a mild adjuvant
protects against low-dose i.v. LCMV challenge (1) but fails
to protect against lethal choriomeningitis (2a). In
contrast, immunization with recombinant vaccinia virus expressing LCMV
glycoprotein induces a strong primary immune response with clearly
detectable ex vivo cytotoxicity and long-lasting protection against
LCMV infection (17). Thus, the induction of protective CTL
by DC is more efficient than priming with peptides but less effective
than immunization with recombinant vaccinia virus. However, only 10,000 to 100,000 DC given i.v. can induce an immune status resembling a
memory situation after virus infection: (i) CTLp frequencies are only
slightly lower than those in LCMV long-term memory mice (3),
i.e., between 1 in 105 to 1 in 104; (ii) ex
vivo cytotoxicity is measurable after prolonged incubation of target
cells with effector cells (15); (iii) mice are protected against i.v. LCMV challenge infection by day 4; and (iv) lethal LCMV-specific immunopathology after intracerebral infection is prevented (35).
Transport of antigen from the periphery to organized lymphoid tissues
by APC is crucial for the initiation of an immune response (7,
25). DC are probably the main APC population contributing to this
antigen transport, as they migrate and home to the T-cell areas of
lymphoid tissues, subsequent to antigen uptake in the periphery
(23, 31). To address the contribution of DC migration to
their immunogenicity, we monitored the migration pathway of bmDC and
correlated it with their ability to induce protective immune responses.
As described previously for sDC (4), bmDC home
preferentially to the interfollicular T-cell areas of the spleen. A
smaller proportion is also found in the liver-draining lymph nodes,
probably due to their ability to translocate into the hepatic lymph
system (23). To prime a protective CTL response in the
spleen, only 100 to 1,000 DC had to reach the organ, as determined by
direct intrasplenic injection. Ten times more cells, i.e., 1,000 to
10,000, had to be injected i.v. to achieve a similar priming
efficiency. After s.c. injection, the majority of the bmDC rapidly
migrated to the local lymph nodes (data not shown). However, even after
s.c. injection of 10,000 to 100,000 DC, a minimal number of 100 to
1,000 DC still must have reached the spleen to induce a local CTL
response and to protect against LCMV challenge infection. Thus, the
migratory and homing capacities of DC are of critical importance in
delivering antigens to the lymphoid environment.
Knowledge about peptide turnover and its influence on priming
efficiency is particularly important for peptide- and cell-based vaccination strategies. During maturation, DC acquire the ability to
present MHC class II-restricted peptides over prolonged periods of time
(>100 h) (13). In contrast, MHC class I-peptide complexes generally show a rapid turnover (40) on DC (13).
A comparison of H8-bmDC constitutively expressing GP33 peptide with
exogenously pulsed B6-sDC and B6-bmDC showed that turnover of
peptide-MHC class I complexes did not influence the capacity to induce
antiviral immunity in vivo. These results confirm and further emphasize previous findings (29, 38) that the transport of exogenously loaded antigenic peptides to regional lymphoid tissues by DC is highly
efficient.
One aim of the current DC research is the development of DC-based
vaccination strategies against tumors and infectious agents. In tumor
patients, therapy with autologous DC is possible (18) and
necessary because of the high variability of tumor antigens. Clearly,
however, individual treatment with adoptively transferred, peptide-
and/or protein-pulsed DC is too complicated for general immunization.
Therefore, other application pathways for delivering antigenic
determinants specifically to DC should be considered. Manickan et al.
(28) have shown that targeting DNA vaccines containing
herpes simplex virus DNA encoding antigenic epitopes specifically to DC
offers an efficient way to induce T-helper- and B-cell responses
against this infectious agent. Further promising results come from
experiments with "gene gun" technology, showing that this type of
immunization mediates antiviral protection (45), probably
via in vivo transfection of skin DC, which subsequently express the
foreign protein in regional lymph nodes (14). In conclusion,
vaccination via DC appears to be an effective way to induce
long-lasting protective antiviral immunity.
| |
ACKNOWLEDGMENTS |
We thank Paul Klenerman and Peter Aichele for helpful discussions
and critical reading of the manuscript and Lenka Vlk for expert
technical assistance.
This work was supported by the Swiss National Science Foundation, the
Deutsche Forschungsgemeinschaft (grants to B.L. and S.E.), and the
Kanton Zürich.
| |
FOOTNOTES |
*
Corresponding author. Institute of Experimental
Immunology, Department of Pathology, University of Zürich,
Schmelzbergstr. 12, CH-8091 Zürich, Switzerland. Phone: 41-1-255 2989. Fax: 41-1-255 4420. E-mail:
LudewigB{at}pathol.unizh.ch.
| |
REFERENCES |
| 1.
|
Aichele, P.,
K. Brduscha-Riem,
R. M. Zinkernagel,
H. Hengartner, and H. Pircher.
1995.
T cell priming versus T cell tolerance induced by synthetic peptides.
J. Exp. Med.
182:261-266[Abstract/Free Full Text].
|
| 2.
|
Aichele, P.,
H. Hengartner,
R. M. Zinkernagel, and M. Schulz.
1990.
Antiviral cytotoxic T cell response induced by in vivo priming with a free synthetic peptide.
J. Exp. Med.
171:1815-1820[Abstract/Free Full Text].
|
| 2a.
| Aichele, P. Personal communication.
|
| 3.
|
Assmann-Wischer, U.,
D. Moskophidis,
M. M. Simon, and F. Lehmann-Grube.
1986.
Numbers of cytolytic T lymphocytes (CTL) and CTL precursor cells in spleens of mice acutely infected with lymphocytic choriomeningitis virus.
Med. Microbiol. Immunol.
175:141-143[Medline].
|
| 4.
|
Austyn, J. M.,
J. W. Kupiec Weglinski,
D. F. Hankins, and P. J. Morris.
1988.
Migration patterns of dendritic cells in the mouse. Homing to T cell-dependent areas of spleen, and binding within marginal zone.
J. Exp. Med.
167:646-651[Abstract/Free Full Text].
|
| 5.
|
Bachmann, M. F., and T. M. Kundig.
1994.
In vivo versus in vitro assays for assessment of T- and B-cell function.
Curr. Opin. Immunol.
6:320-326[Medline].
|
| 6.
|
Bachmann, M. F.,
T. M. Kundig,
H. Hengartner, and R. M. Zinkernagel.
1997.
Protection against immunopathological consequences of a viral infection by activated but not resting cytotoxic T cells: T cell memory without "memory T cells"?
Proc. Natl. Acad. Sci. USA
94:640-645[Abstract/Free Full Text].
|
| 7.
| Barker, C. F., and R. E. Billingham.
1967. The role of regional lymphatics in the skin homograft response.
Transplantation 5(Suppl.):962-966.
|
| 8.
|
Battegay, M.,
S. Cooper,
A. Althage,
J. Banziger,
H. Hengartner, and R. M. Zinkernagel.
1991.
Quantification of lymphocytic choriomeningitis virus with an immunological focus assay in 24- or 96-well plates.
J. Virol. Methods
33:191-198[Medline].
|
| 9.
|
Boczkowski, D.,
S. K. Nair,
D. Snyder, and E. Gilboa.
1996.
Dendritic cells pulsed with RNA are potent antigen-presenting cells in vitro and in vivo.
J. Exp. Med.
184:465-472[Abstract/Free Full Text].
|
| 10.
|
Bohm, W.,
R. Schirmbeck,
A. Elbe,
K. Melber,
D. Diminky,
G. Kraal,
N. van Rooijen,
Y. Barenholz, and J. Reimann.
1995.
Exogenous hepatitis B surface antigen particles processed by dendritic cells or macrophages prime murine MHC class I-restricted cytotoxic T lymphocytes in vivo.
J. Immunol.
155:3313-3321[Abstract].
|
| 11.
|
Borrow, P.,
C. F. Evans, and M. B. Oldstone.
1995.
Virus-induced immunosuppression: immune system-mediated destruction of virus-infected dendritic cells results in generalized immune suppression.
J. Virol.
69:1059-1070[Abstract].
|
| 12.
|
Castelmur, I.,
C. DiPaolo,
M. F. Bachmann,
H. Hengartner,
R. M. Zinkernagel, and T. M. Kundig.
1993.
Comparison of the sensitivity of in vivo and in vitro assays for detection of antiviral cytotoxic T cell activity.
Cell. Immunol.
151:460-466[Medline].
|
| 13.
|
Cella, M.,
A. Engering,
V. Pinet,
J. Pieters, and A. Lanzavecchia.
1997.
Inflammatory stimuli induce accumulation of MHC class II complexes on dendritic cells.
Nature
388:782-787[Medline].
|
| 14.
|
Condon, C.,
S. C. Watkins,
C. M. Celluzi,
K. Thompson, and L. D. Falo.
1996.
DNA-based immunization by in vivo transfection of dendritic cells.
Nat. Med.
2:1122-1128[Medline].
|
| 14a.
| Ehl, S., et al. Submitted for publication.
|
| 15.
| Ehl, S., P. Klenerman, P. Aichele, H. Hengartner, and
R. M. Zinkernagel. A functional and kinetic comparison of
antiviral effector and memory CTL populations in vivo and in vitro.
Eur. J. Immunol., in press.
|
| 16.
|
Flamand, V.,
T. Sornasse,
K. Thielemans,
C. Demanet,
M. Bakkus,
H. Bazin,
F. Tielemans,
O. Leo,
J. Urbain, and M. Moser.
1994.
Murine dendritic cells pulsed in vitro with tumor antigen induce tumor resistance in vivo.
Eur. J. Immunol.
24:605-610[Medline].
|
| 17.
|
Hany, M.,
S. Oehen,
M. Schulz,
H. Hengartner,
M. Mackett,
D. H. Bishop,
H. Overton, and R. M. Zinkernagel.
1989.
Anti-viral protection and prevention of lymphocytic choriomeningitis or of the local footpad swelling reaction in mice by immunization with vaccinia-recombinant virus expressing LCMV-WE nucleoprotein or glycoprotein.
Eur. J. Immunol.
19:417-424[Medline].
|
| 18.
|
Hsu, F. J.,
C. Benike,
F. Fagnoni,
T. M. Liles,
D. Czerwinski,
B. Taidi,
E. G. Engleman, and R. Levy.
1996.
Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells.
Nat. Med.
2:52-58[Medline].
|
| 19.
|
Inaba, K.,
M. Inaba,
M. Naito, and R. M. Steinman.
1993.
Dendritic cell progenitors phagocytose particulates, including bacillus Calmette-Guerin organisms, and sensitize mice to mycobacterial antigens in vivo.
J. Exp. Med.
178:479-488[Abstract/Free Full Text].
|
| 20.
|
Inaba, K.,
M. Inaba,
N. Romani,
H. Aya,
M. Deguchi,
S. Ikehara,
S. Muramatsu, and R. M. Steinman.
1992.
Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor.
J. Exp. Med.
176:1693-1702[Abstract/Free Full Text].
|
| 21.
|
Kagi, D.,
B. Ledermann,
K. Burki,
P. Seiler,
B. Odermatt,
K. J. Olsen,
E. R. Podack,
R. M. Zinkernagel, and H. Hengartner.
1994.
Cytotoxicity mediated by T cells and natural killer cells is greatly impaired in perforin-deficient mice.
Nature
369:31-37[Medline].
|
| 22.
|
Kast, W. M.,
C. J. Boog,
B. O. Roep,
A. C. Voordouw, and C. J. Melief.
1988.
Failure or success in the restoration of virus-specific cytotoxic T lymphocyte response defects by dendritic cells.
J. Immunol.
140:3186-3193[Abstract].
|
| 23.
|
Kudo, S.,
K. Matsuno,
T. Ezaki, and M. Ogawa.
1997.
A novel migration pathway for rat dendritic cells from the blood: hepatic sinusoid-lymph translocation.
J. Exp. Med.
185:777-784[Abstract/Free Full Text].
|
| 24.
|
Kundig, T. M.,
M. F. Bachmann,
S. Oehen,
U. W. Hoffmann,
J. J. L. Simard,
C. P. Kalberer,
H. Pircher,
P. S. Ohashi,
H. Hengartner, and R. M. Zinkernagel.
1996.
On the role of antigen in maintaining cytotoxic T-cell memory.
Proc. Natl. Acad. Sci. USA
93:9716-9723[Abstract/Free Full Text].
|
| 25.
|
Lafferty, K. J.,
A. Bootes,
G. Dart, and D. W. Talmage.
1976.
Effect of organ culture on the survival of thyroid allografts in mice.
Transplantation
22:138-149[Medline].
|
| 26.
|
Macatonia, S. E.,
S. Patterson, and S. C. Knight.
1991.
Primary proliferative and cytotoxic T-cell responses to HIV induced in vitro by human dendritic cells.
Immunology
74:399-406[Medline].
|
| 27.
|
Macatonia, S. E.,
P. M. Taylor,
S. C. Knight, and B. A. Askonas.
1989.
Primary stimulation by dendritic cells induces antiviral proliferative and cytotoxic T cell responses in vitro.
J. Exp. Med.
169:1255-1264[Abstract/Free Full Text].
|
| 28.
|
Manickan, E.,
S. Kanangat,
R. J. Rouse,
Z. Yu, and B. T. Rouse.
1997.
Enhancement of immune response to naked DNA vaccine by immunization with transfected dendritic cells.
J. Leukocyte Biol.
61:125-132[Abstract].
|
| 29.
|
Mayordomo, J. I.,
T. Zorina,
W. J. Storkus,
L. Zitvogel,
C. Celluzzi,
L. D. Falo,
C. J. Melief,
S. T. Ildstad,
W. M. Kast,
A. B. DeLeo, and M. T. Lotze.
1995.
Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.
Nat. Med.
1:1297-1302[Medline].
|
| 30.
|
Mbow, M. L.,
N. Zeidner,
N. Panella,
R. G. Titus, and J. Piesman.
1997.
Borrelia burgdorferi-pulsed dendritic cells induce a protective immune response against tick-transmitted spirochetes.
Infect. Immun.
65:3386-3390[Abstract].
|
| 31.
|
Moll, H.
1993.
Epidermal Langerhans cells are critical for immunoregulation of cutaneous leishmaniasis.
Immunol. Today
14:383-387[Medline].
|
| 32.
|
Moskophidis, D.,
U. Assmann-Wischer,
M. M. Simon, and F. Lehmann-Grube.
1987.
The immune response of the mouse to lymphocytic choriomeningitis virus. V. High numbers of cytolytic T lymphocytes are generated in the spleen during acute infection.
Eur. J. Immunol.
17:937-942[Medline].
|
| 33.
|
Oehen, S.,
K. Brduscha-Riem,
A. Oxenius, and B. Odermatt.
1997.
A simple method for evaluating the rejection of grafted spleen cells by flow cytometry and tracing adoptively transferred cells by light microscopy.
J. Immunol. Methods
207:33-42[Medline].
|
| 34.
|
Oehen, S.,
H. Hengartner, and R. M. Zinkernagel.
1991.
Vaccination for disease.
Science
251:195-198[Abstract/Free Full Text].
|
| 35.
|
Oehen, S.,
H. Waldner,
T. M. Kundig,
H. Hengartner, and R. M. Zinkernagel.
1992.
Antivirally protective cytotoxic T cell memory to lymphocytic choriomeningitis virus is governed by persisting antigen.
J. Exp. Med.
176:1273-1281[Abstract/Free Full Text].
|
| 36.
|
Pircher, H.,
K. Burki,
R. Lang,
H. Hengartner, and R. M. Zinkernagel.
1989.
Tolerance induction in double specific T-cell receptor transgenic mice varies with antigen.
Nature
342:559-561[Medline].
|
| 37.
|
Pircher, H.,
D. Moskophidis,
U. Rohrer,
K. Burki,
H. Hengartner, and R. M. Zinkernagel.
1990.
Viral escape by selection of cytotoxic T cell-resistant virus variants in vivo.
Nature
346:629-633[Medline].
|
| 38.
|
Porgador, A., and E. Gilboa.
1995.
Bone marrow-generated dendritic cells pulsed with a class I-restricted peptide are potent inducers of cytotoxic T lymphocytes.
J. Exp. Med.
182:255-260[Abstract/Free Full Text].
|
| 39.
|
Porgador, A.,
D. Snyder, and E. Gilboa.
1996.
Induction of antitumor immunity using bone marrow-generated dendritic cells.
J. Immunol.
156:2918-2926[Abstract].
|
| 40.
|
Reid, P. A., and C. Watts.
1990.
Cycling of cell-surface MHC glycoproteins through primaquine-sensitive intracellular compartments.
Nature
346:655-657[Medline].
|
| 41.
|
Schulz, M.,
R. M. Zinkernagel, and H. Hengartner.
1991.
Peptide-induced antiviral protection by cytotoxic T cells.
Proc. Natl. Acad. Sci. USA
88:991-993[Abstract/Free Full Text].
|
| 42.
|
Steinman, R. M.
1991.
The dendritic cell system and its role in immunogenicity.
Annu. Rev. Immunol.
9:271-296[Medline].
|
| 43.
|
Takahashi, H.,
Y. Nakagawa,
K. Yokomuro, and J. A. Berzofsky.
1993.
Induction of CD8+ cytotoxic T lymphocytes by immunization with syngeneic irradiated HIV-1 envelope derived peptide-pulsed dendritic cells.
Int. Immunol.
5:849-857[Abstract/Free Full Text].
|
| 44.
|
Taswell, C.
1981.
Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis.
J. Immunol.
126:1614-1619[Abstract].
|
| 45.
|
Zarozinski, C. C.,
E. F. Fynan,
L. K. Selin,
H. L. Robinson, and R. M. Welsh.
1995.
Protective CTL-dependent immunity and enhanced immunopathology in mice immunized by particle bombardment with DNA encoding an internal virion protein.
J. Immunol.
154:4010-4017[Abstract].
|
| 46.
|
Zitvogel, L.,
J. I. Mayordomo,
T. Tjandrawan,
A. B. DeLeo,
M. R. Clarke,
M. T. Lotze, and W. J. Storkus.
1996.
Therapy of murine tumors with tumor peptide-pulsed dendritic cells: dependence on T cells, B7 costimulation, and T helper cell 1-associated cytokines.
J. Exp. Med.
183:87-97[Abstract/Free Full Text].
|
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Copyright © 1998, American Society for Microbiology. All rights reserved.
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-
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-
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[Full Text]
-
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(2002). Preferential Infection of Immature Dendritic Cells and B Cells by Mouse Mammary Tumor Virus. J. Immunol.
168: 3470-3476
[Abstract]
[Full Text]
-
Bacci, A., Montagnoli, C., Perruccio, K., Bozza, S., Gaziano, R., Pitzurra, L., Velardi, A., d'Ostiani, C. F., Cutler, J. E., Romani, L.
(2002). Dendritic Cells Pulsed with Fungal RNA Induce Protective Immunity to Candida albicans in Hematopoietic Transplantation. J. Immunol.
168: 2904-2913
[Abstract]
[Full Text]
-
Voehringer, D., Blaser, C., Brawand, P., Raulet, D. H., Hanke, T., Pircher, H.
(2001). Viral Infections Induce Abundant Numbers of Senescent CD8 T Cells. J. Immunol.
167: 4838-4843
[Abstract]
[Full Text]
-
Lambrecht, B.N., Prins, J-;B., Hoogsteden, H.C.
(2001). Lung dendritic cells and host immunity to infection. Eur Respir J
18: 692-704
[Abstract]
[Full Text]
-
Lisziewicz, J., Gabrilovich, D. I., Varga, G., Xu, J., Greenberg, P. D., Arya, S. K., Bosch, M., Behr, J.-P., Lori, F.
(2001). Induction of Potent Human Immunodeficiency Virus Type 1-Specific T-Cell-Restricted Immunity by Genetically Modified Dendritic Cells. J. Virol.
75: 7621-7628
[Abstract]
[Full Text]
-
Worgall, S., Kikuchi, T., Singh, R., Martushova, K., Lande, L., Crystal, R. G.
(2001). Protection against Pulmonary Infection with Pseudomonas aeruginosa following Immunization with P. aeruginosa-Pulsed Dendritic Cells. Infect. Immun.
69: 4521-4527
[Abstract]
[Full Text]
-
Mikloska, Z., Bosnjak, L., Cunningham, A. L.
(2001). Immature Monocyte-Derived Dendritic Cells Are Productively Infected with Herpes Simplex Virus Type 1. J. Virol.
75: 5958-5964
[Abstract]
[Full Text]
-
Rudolf, M. P., Fausch, S. C., Da Silva, D. M., Kast, W. M.
(2001). Human Dendritic Cells Are Activated by Chimeric Human Papillomavirus Type-16 Virus-Like Particles and Induce Epitope-Specific Human T Cell Responses In Vitro. J. Immunol.
166: 5917-5924
[Abstract]
[Full Text]
-
Ludewig, B., McCoy, K., Pericin, M., Ochsenbein, A. F., Dumrese, T., Odermatt, B., Toes, R. E. M., Melief, C. J. M., Hengartner, H., Zinkernagel, R. M.
(2001). Rapid Peptide Turnover and Inefficient Presentation of Exogenous Antigen Critically Limit the Activation of Self-Reactive CTL by Dendritic Cells. J. Immunol.
166: 3678-3687
[Abstract]
[Full Text]
-
Peggs, K., Verfuerth, S., Mackinnon, S.
(2001). Induction of cytomegalovirus (CMV)-specific T-cell responses using dendritic cells pulsed with CMV antigen: a novel culture system free of live CMV virions. Blood
97: 994-1000
[Abstract]
[Full Text]
-
Ludewig, B., Freigang, S., Jäggi, M., Kurrer, M. O., Pei, Y.-C., Vlk, L., Odermatt, B., Zinkernagel, R. M., Hengartner, H.
(2000). Linking immune-mediated arterial inflammation and cholesterol-induced atherosclerosis in a transgenic mouse model. Proc. Natl. Acad. Sci. USA
10.1073/pnas.220427097v1
[Abstract]
[Full Text]
-
Chang, C.-C. J., Wright, A., Punnonen, J.
(2000). Monocyte-Derived CD1a+ and CD1a- Dendritic Cell Subsets Differ in Their Cytokine Production Profiles, Susceptibilities to Transfection, and Capacities to Direct Th Cell Differentiation. J. Immunol.
165: 3584-3591
[Abstract]
[Full Text]
-
Kruse, M., Rosorius, O., Krätzer, F., Stelz, G., Kuhnt, C., Schuler, G., Hauber, J., Steinkasserer, A.
(2000). Mature Dendritic Cells Infected with Herpes Simplex Virus Type 1 Exhibit Inhibited T-Cell Stimulatory Capacity. J. Virol.
74: 7127-7136
[Abstract]
[Full Text]
-
Ludewig, B., Ochsenbein, A. F., Odermatt, B., Paulin, D., Hengartner, H., Zinkernagel, R. M.
(2000). Immunotherapy with Dendritic Cells Directed against Tumor Antigens Shared with Normal Host Cells Results in Severe Autoimmune Disease. JEM
191: 795-804
[Abstract]
[Full Text]
-
Liu, L., Usherwood, E. J., Blackman, M. A., Woodland, D. L.
(1999). T-Cell Vaccination Alters the Course of Murine Herpesvirus 68 Infection and the Establishment of Viral Latency in Mice. J. Virol.
73: 9849-9857
[Abstract]
[Full Text]
-
Ahuja, S. S., Reddick, R. L., Sato, N., Montalbo, E., Kostecki, V., Zhao, W., Dolan, M. J., Melby, P. C., Ahuja, S. K.
(1999). Dendritic Cell (DC)-Based Anti-Infective Strategies: DCs Engineered to Secrete IL-12 Are a Potent Vaccine in a Murine Model of an Intracellular Infection. J. Immunol.
163: 3890-3897
[Abstract]
[Full Text]
-
Ludewig, B., Oehen, S., Barchiesi, F., Schwendener, R. A., Hengartner, H., Zinkernagel, R. M.
(1999). Protective Antiviral Cytotoxic T Cell Memory Is Most Efficiently Maintained by Restimulation Via Dendritic Cells. J. Immunol.
163: 1839-1844
[Abstract]
[Full Text]
-
Renjifo, X., Letellier, C., Keil, G. M., Ismaili, J., Vanderplasschen, A., Michel, P., Godfroid, J., Walravens, K., Charlier, G., Pastoret, P.-P., Urbain, J., Denis, M., Moser, M., Kerkhofs, P.
(1999). Susceptibility of Bovine Antigen-Presenting Cells to Infection by Bovine Herpesvirus 1 and In Vitro Presentation to T Cells: Two Independent Events. J. Virol.
73: 4840-4846
[Abstract]
[Full Text]
-
Zarling, A. L., Johnson, J. G., Hoffman, R. W., Lee, D. R.
(1999). Induction of Primary Human CD8+ T Lymphocyte Responses In Vitro Using Dendritic Cells. J. Immunol.
162: 5197-5204
[Abstract]
[Full Text]
-
Lu, H., Zhong, G.
(1999). Interleukin-12 Production Is Required for Chlamydial Antigen-Pulsed Dendritic Cells To Induce Protection against Live Chlamydia trachomatis Infection. Infect. Immun.
67: 1763-1769
[Abstract]
[Full Text]
-
Ochsenbein, A. F., Klenerman, P., Karrer, U., Ludewig, B., Pericin, M., Hengartner, H., Zinkernagel, R. M.
(1999). Immune surveillance against a solid tumor fails because of immunological ignorance. Proc. Natl. Acad. Sci. USA
96: 2233-2238
[Abstract]
[Full Text]
-
Maloy, K. J., Burkhart, C., Freer, G., Rulicke, T., Pircher, H., Kono, D. H., Theofilopoulos, A. N., Ludewig, B., Hoffmann-Rohrer, U., Zinkernagel, R. M., Hengartner, H.
(1999). Qualitative and Quantitative Requirements for CD4+ T Cell-Mediated Antiviral Protection. J. Immunol.
162: 2867-2874
[Abstract]
[Full Text]
-
Redchenko, I. V., Rickinson, A. B.
(1999). Accessing Epstein-Barr Virus-Specific T-Cell Memory with Peptide-Loaded Dendritic Cells. J. Virol.
73: 334-342
[Abstract]
[Full Text]
-
Ludewig, B., Odermatt, B., Landmann, S., Hengartner, H., Zinkernagel, R. M.
(1998). Dendritic Cells Induce Autoimmune Diabetes and Maintain Disease via De Novo Formation of Local Lymphoid Tissue. JEM
188: 1493-1501
[Abstract]
[Full Text]
-
Ludewig, B., Freigang, S., Jaggi, M., Kurrer, M. O., Pei, Y.-C., Vlk, L., Odermatt, B., Zinkernagel, R. M., Hengartner, H.
(2000). Linking immune-mediated arterial inflammation and cholesterol-induced atherosclerosis in a transgenic mouse model. Proc. Natl. Acad. Sci. USA
97: 12752-12757
[Abstract]
[Full Text]
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Abstract
Introduction
