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Malaghan Institute of Medical Research, Wellington School of Medicine, Wellington, New Zealand
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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but not IL-4 or IL-5 by
host cells. To further address the role of IL-4 and IL-5 in antitumor
immunity, tumor-specific TCR-transgenic CD8+ T cells were
activated in vitro to generate cytotoxic T (Tc) 1 cells that secrete
high IFN- and no IL-4 or IL-5 or Tc2 cells that secrete IL-4, IL-5,
and some IFN-. Both cell types killed target cells in vitro. Tc1 and
Tc2 cells were adoptively transferred into syngeneic hosts, and their
ability to protect against tumor challenge was compared. Tc1 cells were
able to significantly delay tumor growth, whereas Tc2 cells or Tc2
cells from IFN--/- donors had no effect. This was due
to neither the inability of Tc2 cells to survive in vivo or to migrate
to the tumor site nor their inability to secrete IL-4 and/or IL-5 in
the presence of limiting amounts of anti-CD3. However, IFN-
secretion by Tc2 cells was triggered inefficiently by restimulation
with Ag compared with anti-CD3. We conclude that the ability to
secrete "type 2" cytokines, and cytotoxic ability, have a limited
role in antitumor immune responses mediated by CD8+ T
cells, whereas the capacity to secrete high amounts of IFN- remains
the most critical antitumor effector mechanism in
vivo. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In physiological situations, infiltration of the tumor mass by effector
cells is dependent on recognition of tumor Ags by specific T cells.
Both CD4+ and CD8+ T cells
have been demonstrated to play critical roles in this process
(3). Removal of CD8+ T cells causes
enhanced tumor growth (4); conversely, the adoptive
transfer of antitumor CD8+ T cells results in
delayed tumor growth (5, 6, 7). In addition, immunization
procedures that activate CD8+ T cell immune
responses, e.g., DC vaccination, induce protection against tumor growth
(8, 9, 10). The antitumor effect of
CD8+ T cells is presumably mediated through the
ability of these cells to lyse tumor cells and to secrete cytokines
upon recognition of Ag on tumor cells. CD4+ T
cells are also critical to the antitumor effect (11, 12, 13, 14, 15).
Indeed, due to their capacity to secrete multiple cytokines,
CD4+ helper T cells may have a better ability to
activate antitumor effector mechanisms. Both Th1 cytokines such as
IFN- and TNF-
and Th2 cytokines such as IL-4, IL-5, and GM-CSF
can contribute to the antitumor effect (16).
It has become apparent in recent years that CD8+
cytotoxic T (Tc) cells are also heterogeneous in their ability to
secrete cytokines. Similarly to Th cells, Tc cells can also be
subdivided on the basis of the cytokines they secrete into Tc1, which
secrete IFN- and TNF-, and Tc2 which secrete IL-4,
IL-5 and IL-10 (17, 18, 19). There is variability in the
degree to which Tc2 cells can be polarized away from the default Tc1
phenotype of IFN- secretion and cytotoxic activity
(17, 18, 19). In contrast, acquisition of the capacity to
secrete IL-4 and IL-5 appears to be induced easily and reproducibly
through in vitro culture in the presence of IL-4 (17, 18, 19).
We took advantage of this observation to investigate whether Tc cells
with a multiple cytokine secretion capacity also have an improved
ability to mediate tumor rejection. We report here that whereas
tumor-specific Tc2 cells have little or no effect on tumor growth,
tumor-specific Tc1 cells have a strong antitumor effect.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Female and male C57BL/6J mice were originally obtained from The
Jackson Laboratory (Bar Harbor, ME). The "line 318" mouse strain,
transgenic for a TCR specific for H-2Db +
fragment 33–41 of the lymphocytic choriomeningitis virus (LCMV)
glycoprotein (LCMV33–41) was kindly provided by
Dr. H. Pircher (Institute of Medical Microbiology, University of
Freiburg, Freiburg, Germany) (20). IL-4 and IL-5 gene
knockout mice (21, 22) were obtained from Dr. M. Kopf
(Basel Institute of Immunology, Basel, Switzerland). IFN- gene
knockout mice (23) were purchased from The Jackson
Laboratory and were backcrossed twice to line 318 mice to obtain
IFN--/- 318 progeny for experimental
use.
All animals were maintained by brother x sister mating at the Biomedical Research Unit of the Wellington School of Medicine (Wellington, New Zealand). All in vivo experiments were approved by the Wellington School of Medicine Ethics Committee and performed according to institutional guidelines.
Tumor cell lines and in vitro culture medium
The tumor cell line LL-LCMV is a derivative of the Lewis lung carcinoma LLTC (C57BL/6J, H-2b) that has been modified to express a minigene encoding LCMV33–41 under the control of a CMV promoter (24). Unless otherwise stated, all cultures were in IMDM (Life Technologies, Auckland, New Zealand) containing 2 mM glutamine, 1% penicillin-streptomycin, 5 x 10-5 M 2-ME (all from Sigma, St. Louis, MO), and 5% FCS (Life Technologies). LL-LCMV tumor cells were maintained in culture medium containing 0.5 mg/ml G418 (Life Technologies). The synthetic peptide LCMV33–41 (KAVYNFATM) was obtained from Chiron Mimotopes (Clayton, Australia).
DC immunization and tumor challenge experiments
Bone marrow cells from C57BL/6J mice were cultured in medium containing 20 ng/ml IL-4 and 20 ng/ml GM-CSF as described (25). DC were harvested on day 7, loaded with peptide by incubation in medium containing 10 µM LCMV33–41 for 2 h at 37°C, and washed three times in IMDM before in vivo injection. For in vivo tumor challenge experiments, 1 x 105 DC that had been loaded with peptide or left untreated were injected s.c. into the left flank on day 0. Mice were challenged with 5 x 105 tumor cells injected into the opposite flank on day 7. Mean tumor size was calculated as the mean product of bisecting tumor diameters as described (24).
In vitro generation of Tc1 and Tc2 cells and adoptive transfer
Single-cell suspensions from spleen and lymph nodes of line 318
mice or IFN--/- 318 mice were washed in
incomplete medium and resuspended at 2 x
106/ml. Cells were incubated for 2 h at
37°C to deplete adherent cells, and CD4+ T
lymphocytes and B lymphocytes were removed by Ab treatment
(GK1.5-biotin) and magnetic depletion using Dynabeads conjugated to
streptavidin or rat anti-mouse IgG (Dynal, Victoria, Australia).
Recovered cells were 90–98% CD8+ as determined
by FACS staining.
The purified CD8+ T cells were activated in
six-well plates (Falcon, Oxnard, CA) that were coated overnight at
4°C with 5 µg/ml mouse anti-Armenian hamster IgG and 2 µg/ml
mouse anti-Syrian hamster IgG (both from BD PharMingen, San Diego,
CA) in 0.15 M NaCl, 0.015 M boric acid (pH 8.5). Plates were washed
three times in PBS before purified anti-CD3 Ab (145-2C11) was added
in PBS for 2 h at 37°C; then plates were washed three times in
PBS. For Tc1 cell generation, naive CD8+ T cells
(1 x 106/ml) were added to the plates with
10 U/ml recombinant human IL-2, 10 ng/ml IL-6, and 2% 37.51 hybridoma
supernatant (anti-CD28). For Tc2 cell generation, IL-4-containing
supernatant was also added at 2000 U/ml. Cultures were maintained for 5
days, with replacement of medium and cytokines on days 2, 3, and 4. On
day 5, cells were harvested and cultured in fresh six-well plates in
medium containing 100 U/ml recombinant human IL-2 for 48 h, with
replacement of IL-2 after 24 h. Cells were washed in medium before
use and were at least 75%
V2+V
8+ by FACS
analysis.
C57BL/6J mice received up to 7 x 106 in
vitro-generated
V2+V8+ Tc1 or Tc2
effector CD8+ T cells by i.v. injection in the
lateral tail vein. Tumor challenge took place 24 h later.
Cytotoxicity assay
The cytotoxic activity of Tc1 and Tc2 cells was determined using the just another method (JAM) test (26). Briefly, 5000 [3H]thymidine-labeled EL4 cells were incubated in 96-well U-bottom plates with or without 0.01 µM LCMV33–41 peptide for 1 h. Cultured CD8+ effector T cells were added at different ratios and plates incubated for a further 3.5 h. Percent cytotoxicity was calculated from the mean of triplicate wells as described (24).
Cytokine secretion by Tc cells
Tc1 and Tc2 cells were restimulated at 1 x
106/ml in 96-well plates (Nunc, Roskilde,
Denmark) coated with 10 µg/ml anti-CD3 or in the presence of
1 x 105 irradiated spleen cells and 10 µM
LCMV33–41 in complete medium for 48 h.
Control cultures received no spleen cells. Cytokine-containing
supernatants were harvested and analyzed by cytokine-specific ELISA
using the appropriate Ab pairs (TRFK5 and TRFK4-biotin for IL-5; 11B11
and BVD6-24G2-biotin for IL-4; AN18 and XMGD6-biotin for IFN-) and
mouse anti-IL-10 mAbs (BD Labware, Mountain View, CA) as described
(27). Standard curves were generated using recombinant
cytokines from hybridoma cultures: IL-4 (25 ng/ml), IL-5 (300 ng/ml),
and IFN- (60 ng/ml) or IL-10 (5 ng/ml; BD Labware). Cytokine
secretion by T cells or stimulator cells in medium alone was
undetectable.
Flow cytometry
Anti-FcRII (2.4G2), anti-CD8-FITC (2.43), and
anti-V8-biotin (KJ16) mAb were affinity purified from tissue
culture supernatants using protein G-Sepharose (Pharmacia Biotech,
Uppsala, Sweden) and conjugated to FITC or biotin. Anti-V2-PE was
obtained from BD PharMingen. Cells were stained in PBS containing 2%
FCS and 0.01% sodium azide as described (25) and analyzed
on a FACSort (BD Biosciences, Mountain View, CA). Live cells were gated
on the basis of forward and side scatter profiles.
CFSE labeling and adoptive transfer
In vitro-activated cells were washed in PBS and resuspended at 2 x 107/ml in PBS. CFSE (Molecular Probes, Eugene, OR) was used at 1.2 µM in PBS and added to cells for 8 min at room temperature. For migration and survival studies, lymph nodes, spleen, or s.c. tissue corresponding to the tumor site were taken from animals at the indicated times after tumor challenge. Single-cell suspensions were obtained and analyzed by flow cytometry.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, but
not IL-4 or IL-5
Immunization with DC loaded with tumor peptide is an effective
method to induce antitumor CD8+ T cell responses
(8, 9, 28). We have previously shown that immunization
with DC loaded with LCMV33–41 induces protective
immunity against challenge with the
LCMV33–41-expressing tumor cell line LL-LCMV
(24). We used this experimental system to investigate the
role of T cell-derived cytokines in the antitumor immune response.
C57BL/6J control mice and IFN--/-,
IL-4-/-, and IL-5-/-
mice were immunized with either C57BL/6J DC or DC loaded with
LCMV33–41 peptide or were left nonimmunized. One
week later, all mice were challenged by s.c. injection of LL-LCMV tumor
cells. All mice that received no DC immunization or that were immunized
with DC only developed tumors that appeared and progressed with similar
kinetics. As previously reported, C57BL/6J mice immunized with
peptide-loaded DC showed delayed growth of LL-LCMV tumors as compared
with mice injected with DC alone or nonimmunized mice (Fig. 1
and Ref. 24). In contrast,
no delay in tumor growth could be demonstrated in
IFN--/- mice that had been immunized with
peptide-loaded DC (Fig. 1), indicating that IFN- secretion by host
cells is essential for the antitumor effect induced by DC immunization.
Similar results have also been reported by Zitvogel et al.
(11). Delayed tumor growth was observed in
IL-4-/- and IL-5-/-
recipient mice (Fig. 1), indicating that IL-4 and IL-5 were not
essential to the antitumor immune response induced by DC immunization.
In three separate experiments, tumor growth was always delayed further
in IL-4-/- and IL-5 -/-
mice than in C57BL/6J mice, suggesting that these cytokines had an
inhibitory effect on the antitumor immune response.
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The apparent lack of involvement of IL-4 or IL-5 in DC-induced
antitumor immune responses could indicate that IL-4- or IL-5-secreting
T cells had not been induced by the immunization protocol. Furthermore,
it was unclear from the DC immunization experiments whether IFN-
played a role during the priming of antitumor T cells, or in the
effector phase, or both. To resolve these issues, we examined the
antitumor effect of T cells activated in vitro.
T cells were obtained from line 318 mice, which carry a transgenic TCR
specific for Db plus
LCMV33–41. The same Ag is also expressed on the
LL-LCMV tumor cell line. We have previously shown that T cells from
line 318 mice can proliferate in the presence of tumor cells in vitro
and that their activation in vivo correlates with antitumor immunity
(24, 29). Tc1 cells were generated by activating purified
line 318 CD8+ T cells in vitro with cross-linked
anti-CD3 and anti-CD28 in the presence of IL-2 and IL-6; Tc2
cells were generated using identical conditions, but with the addition
of IL-4. These conditions were chosen after extensive preliminary
experiments in which the effect of cytokines such as IL-12 and
neutralizing Abs to IL-4 and IFN- were tested and found to have
little effect on the phenotype of the resulting cells (R. A. Kemp,
manuscript in preparation). After 7 days of culture, T cells were
harvested and restimulated with anti-CD3 to induce cytokine
production. Fig. 2 shows that Tc1 cells
generated from these cultures secreted IFN-, but no IL-4 or IL-5,
and only small amounts of IL-10. In contrast, Tc2 cells secreted
considerable quantities of IL-4, IL-5, and IL-10 but less IFN- than
Tc1 cells. No culture conditions generating Tc cells that did not
secrete IFN- were identified. Thus, to evaluate the role of IFN-
in tumor immunity more directly, Tc2 cells were generated from line 318
mice that had been backcrossed to IFN--/-
mice. As shown in Fig. 2, Tc2 cells from these mice produced no IFN-
but secreted larger amounts of IL-4, IL-5, and IL-10 than did Tc2 from
IFN-+/+ mice, suggesting that IFN- may have
an inhibitory effect on the development of cells producing type 2
cytokines. In summary, we generated three populations of effector
CD8+ T cells, each with distinct cytokine
secretion profiles: a "classical" Tc1 population, secreting IFN-
but no IL-4 or IL-5; a Tc2 population, secreting IL-4, IL-5, and some
IFN-; and an IFN--/- Tc2 population
secreting IL-4 and IL-5 but no IFN-.
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-/- Tc2 cells from in vitro culture. EL4
cells that had been left unpulsed were not killed by any of the cell
types (Fig. 3). In contrast, EL4 cells
loaded with specific Ag were killed by each of the three
CD8+ T cell populations to a similar extent,
indicating that in vitro cytotoxicity was independent of the cytokine
secretion potential.
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-/- Tc2 cells, can
transfer antitumor protective immunity in vivo
To evaluate the antitumor potential of in vitro-activated Tc1,
Tc2, and IFN--/- Tc2 cells, we adoptively
transferred each of these cell types into C57BL/6J mice. Control mice
received no adoptive transfer of T cells. All mice were challenged the
next day with LL-LCMV tumor cells, and tumor growth was measured twice
weekly thereafter. Fig. 4A
shows that mice that had received adoptive transfer of Tc1 cells had
delayed tumor growth compared with control mice. In contrast, mice that
had received Tc2 cells or IFN--/- Tc2 cells
showed no delay in tumor growth. Combined data from several experiments
are shown in Fig. 4B. These results suggest that the
production of IL-4 and IL-5 by CD8+ T cells was
not sufficient to mediate an antitumor effect. Surprisingly, Tc2 cells
also failed to cause a significant delay in tumor growth (Fig. 4A) despite their in vitro cytotoxic activity and their
ability to produce some IFN- as well as IL-4 and IL-5.
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The results shown in Fig. 4
indicate that, unlike Tc1 cells, Tc2
cells were ineffective in controlling tumor growth. To address whether
Tc1 and Tc2 cells differed with respect to their in vivo survival
potential, numbers of CFSE-labeled Tc1 or Tc2 cells in the spleen,
lymph node, and tumors were evaluated by FACS analysis at different
times after adoptive transfer. As shown in Table I, the numbers of CFSE-labeled Tc1 or Tc2
cells recovered from each recipient mouse tissue were similar and
remained relatively constant until day 13 after transfer. The numbers
of CFSE+ cells recovered at the tumor injection
site were similar for Tc1 and Tc2 cells, indicating that both cell
populations had the capacity to migrate to the tumor site. We conclude
that the poor ability of Tc2 cells to mediate tumor protection was not
due to their failure to survive in vivo or to reach the tumor
site.
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We hypothesized that the inability of Tc2 cells to mediate an
antitumor effect could be due to the requirement for a higher degree of
TCR cross-linking to secrete cytokines. To test this hypothesis, Tc1,
Tc2, and IFN--/- Tc2 cells were stimulated
in vitro in the presence of increasing concentrations of plate-bound
anti-CD3, and their secretion of IFN-, IL-4, and IL-5 was
compared. Fig. 5 shows that IFN- could
be detected when Tc1 or Tc2 cells were cultured on plates coated with
0.1 µg/ml anti-CD3. Progressively higher IFN- secretion could
be demonstrated when the amount of anti-CD3 was increased. As
expected, IFN- secretion by Tc1 cells was higher than for Tc2 cells
at each anti-CD3 concentration tested, and no IFN- could be
demonstrated in cultures of IFN- -/-
Tc2 cells. Secretion of IL-4 and IL-5 by Tc2 cells was also detected
upon stimulation with 0.1 µg/ml anti-CD3, and at even lower
concentrations in the case of IFN- -/- Tc2
cells. Together, these results do not support the possibility that Tc2
cells or IFN--/- Tc2 cells may require
higher degrees of TCR cross-linking than Tc1 cells to secrete
cytokines.
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by Tc1 cells could be obtained with both types of
stimulation. As shown in Fig. 6, Tc1
cells secreted comparable amounts of IFN- whether restimulated with
anti-CD3 or with LCMV33–41. In contrast, Tc2
cells secreted 
14-fold less IFN- in the presence of
LCMV33–41 than anti-CD3. This finding was
reproducible over many experiments; on average, the amount of IFN-
secreted by Tc2 cells was 37 ± 9% of the amount produced by Tc1
after anti-CD3 stimulation and 8 ± 3% for stimulation with
Ag and APC. Secretion of IL-4 and IL-5 by Tc2 cells was also compared
in these experiments. As shown in Fig. 6, there was only a 2- to 3-fold
decrease in the amount of IL-4 and IL-5 secreted by Tc2 cells after Ag
stimulation, as compared with anti-CD3 stimulation. Addition of
peptide alone resulted in undetectable amounts of secreted
cytokine.
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after Ag-specific
stimulation in vivo. |
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, whereas the secretion of
other effector cytokines such as IL-4 or IL-5 did not appear to
contribute to the antitumor effect. We also observed that secretion of
IFN- by Tc2 cells was not sufficient to mediate protective antitumor
immunity in vivo.
The lack of antitumor effect of Tc2 cells could not be due to poor
survival in vivo, because similar numbers of Tc1 and Tc2 cells were
recovered from recipient mice (Table I). In support of our observation,
data by other authors also show that T cells activated in the presence
of IL-4, and presumably endowed of a Tc2 cytokine secretion pattern,
survive and maintain their cytotoxic activity after in vivo transfer
(30, 31).
"Type 2" cytokines, such as IL-4 or IL-5, have been reported to
mediate a powerful antitumor effect in vivo (1, 2, 16, 32, 33). The adoptive transfer of tumor-specific Tc2 cells with
cytokine secretion profiles similar to the Tc2 cells used here had
clear effects on tumor growth in vivo (34). In addition,
tumor cells producing IL-4 were readily rejected through a
Th2/Tc2-dependent mechanism (32, 33, 35, 36). We were
surprised to find that, in our system, Tc2 cells and
IFN--/- Tc2 cells did not appear to have an
antitumor effect in vivo, despite their ability to secrete considerable
amounts of IL-4 and IL-5 and lyse tumor target cells in vitro. It is
possible, although unlikely, that the type of tumor used in our
experiments is especially resistant to the action of type 2 cytokines.
Alternatively, the amount of Ag expressed by our tumor cell line may be
insufficient to attract a response from Tc2 cells. In support of this
possibility, the LL-LCMV tumor cell line used in our studies was
selected to express very low amounts of the
LCMV33–41 epitope recognized by the adoptively
transferred Tc cells (24), because this was considered to
represent a more physiological model of tumor antigenicity. In
contrast, other studies have used tumor cells known to express high
amounts of Ag (34, 37) or for which the expression of
tumor Ags had not been characterized (32, 33).
In the course of our experiments, we detected no evidence that Tc2
cells may require especially high TCR cross-linking to secrete
cytokines. Tc1 and Tc2 cells required stimulation with similar
concentrations of anti-CD3 to secrete their typical cytokine
pattern, whereas cytokine secretion by
IFN--/- Tc2 was detected at even lower
amounts of anti-CD3. Because Tc1 cells were clearly able to mediate
an antitumor effect, and this was presumably due to IFN- secretion,
we propose that the conditions at the tumor site were likely to be
conducive to cytokine secretion from not only Tc1 cells but also all
types of Tc cells. However, it is not known whether recognition of
tumor Ags by T cells occurs directly on the tumor cells, as is
generally assumed, or on local APC that have taken up tumor Ag and
present it to T cells. If recognition on local APC was required for any
of the T cells used in this study, it is very unlikely that the low
level of LCMV33–41 epitope expressed by LL-LCMV
would be sufficient to remain stimulatory after cross-presentation by
local APC.
We do not know whether the antitumor effect of adoptively transferred
Tc1 cells was due to their ability to kill tumor cells or to their
ability to secrete cytokines at the tumor site. DC immunization
experiments showed that IFN- secretion was critical to the antitumor
effect (Ref. 11 and Fig. 1); thus, we presume that the
effect of Tc1 cells was mediated, at least in part, via IFN-
secretion. Although IFN- treatment significantly up-regulates MHC
class I expression on LL-LCMV tumor cells in vitro (data not shown), it
does not show a direct effect on tumor cell viability as has been
reported in other systems (38). Thus, a synergistic effect
between IFN- secretion and cytotoxic function may best explain our
observations. Additional Tc1-derived cytokines may also contribute to
the antitumor effect.
The Tc2 cells used in our study acquired the ability to produce IL-4
and IL-5 but also retained the capacity to secrete IFN-. Therefore,
it was unexpected that Tc2 cells had no antitumor effect when, in our
system, IFN- had such a dramatic role in tumor rejection (Fig. 1).
This finding is probably explained by the observation that Tc2 cells
had a much lower capacity to secrete IFN- when stimulated on Ag and
APC than anti-CD3 (Fig. 6) and therefore were probably unable to
secrete sufficiently high amounts of IFN- in the tumor context. The
differential response to Ag or anti-CD3 stimulation was observed
only for Tc2 cells and IFN- and did not extend to Tc1 cells or to
other functions of Tc2 cells such as secretion of type 2 cytokines. We
did not attempt to clarify the basis of this finding, but two
possibilities appear especially likely. The first possibility is that
secretion of higher amounts of the inhibitory cytokines IL-10 by Tc2
cells may regulate APC function in culture. Indeed, IL-10 has been
reported to act in an APC-dependent manner to selectively inhibit
IFN- secretion by CD4+ and
CD8+ T cells, without affecting cytokine
secretion by Th2 clones (39). This effect obviously would
not be observed when Tc2 cells are stimulated with anti-CD3, as was
the case in our experiments. A second possibility is that changes in Ag
recognition may result in the selective loss of some T cells responses,
but not others, by Tc2 cells. A similar situation has been reported for
the recognition of altered peptide ligands by T cell clones
(40). Clearly, further analysis is required to assess
whether secretion of IL-10, or differences in TCR signaling, are at the
basis of our observation. Lastly, the lack of antitumor activity of Tc2
cells might not be due to insufficient production of IFN-, but to an
inhibitory effect of type 2 cytokines on IFN- function at the tumor
site (41, 42).
In conclusion, we propose that the production of IFN- by
tumor-specific CD8+ T cells is critical, although
not sufficient, for antitumor immunity. Production of IL-4 and IL-5 is
less critical, and their importance may depend on the tumor model used.
Thus, our data indicate that antitumor immunotherapies should aim at
inducing maximal Tc1 immune responses, even if this may compromise the
ability to secrete other cytokines.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Franca Ronchese, Malaghan Institute of Medical Research, P.O. Box 7060, Wellington, New Zealand. E-mail address: fronchese{at}malaghan.org.nz
3 Abbreviations used in this paper: DC, dendritic cells; JAM, just another method; Tc, cytotoxic CD8+ T lymphocyte; LCMV, fragment 33–41 of the lymphocytic choriomeningitis virus glycoprotein (LCMV33–41).
Received for publication July 6, 2001. Accepted for publication September 28, 2001.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and tumour necrosis factor have a role in tumour regressions mediated by murine CD8+ tumour-infiltrating lymphocytes. J. Exp. Med. 173:647.. Proc. Natl. Acad. Sci. USA 96:8633. genes. Science 259:1739.1 inhibits antigen-induced lung eosinophilia but not Th2 cell development or recruitment in the lung. Eur. J. Immunol. 27:155.[Medline]
(IFN-) in organ culture of B16 melanoma cells and heart tissue. Int. J. Cancer 42:470.[Medline]
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G. Iezzi, A. Boni, E. Degl'Innocenti, M. Grioni, M. T. S. Bertilaccio, and M. Bellone Type 2 Cytotoxic T Lymphocytes Modulate the Activity of Dendritic Cells Toward Type 2 Immune Responses J. Immunol., August 15, 2006; 177(4): 2131 - 2137. [Abstract] [Full Text] [PDF]
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J. Yang, S. P. Huck, R. S. McHugh, I. F. Hermans, and F. Ronchese Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8+ T cells in vivo PNAS, January 3, 2006; 103(1): 147 - 152. [Abstract] [Full Text] [PDF]
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S. Olver, P. Groves, K. Buttigieg, E. S. Morris, M. L. Janas, A. Kelso, and N. Kienzle Tumor-Derived Interleukin-4 Reduces Tumor Clearance and Deviates the Cytokine and Granzyme Profile of Tumor-Induced CD8+ T Cells Cancer Res., January 1, 2006; 66(1): 571 - 580. [Abstract] [Full Text] [PDF]
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 D. M. Jelley-Gibbs, J. P. Dibble, S. Filipson, L. Haynes, R. A. Kemp, and S. L. Swain Repeated stimulation of CD4 effector T cells can limit their protective function J. Exp. Med., April 4, 2005; 201(7): 1101 - 1112. [Abstract] [Full Text] [PDF]
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N. Kienzle, S. Olver, K. Buttigieg, P. Groves, M. L. Janas, A. Baz, and A. Kelso Progressive Differentiation and Commitment of CD8+ T Cells to a Poorly Cytolytic CD8low Phenotype in the Presence of IL-4 J. Immunol., February 15, 2005; 174(4): 2021 - 2029. [Abstract] [Full Text] [PDF]
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Q. Li, A. L. Carr, E. J. Donald, J. J. Skitzki, R. Okuyama, L. M. Stoolman, and A. E. Chang Synergistic Effects of IL-12 and IL-18 in Skewing Tumor-Reactive T-Cell Responses Towards a Type 1 Pattern Cancer Res., February 1, 2005; 65(3): 1063 - 1070. [Abstract] [Full Text] [PDF]
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 J. M. Roberts, J. Yang, and F. Ronchese IL-4 deficiency does not impair the ability of dendritic cells to initiate CD4+ and CD8+ T cell responses in vivo Int. Immunol., October 1, 2004; 16(10): 1451 - 1458. [Abstract] [Full Text] [PDF]
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R. A. Kemp, T. J. Powell, D. W. Dwyer, and R. W. Dutton Cutting Edge: Regulation of CD8+ T Cell Effector Population Size J. Immunol., September 1, 2004; 173(5): 2923 - 2927. [Abstract] [Full Text] [PDF]
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J. A. Hollenbaugh, J. Reome, M. Dobrzanski, and R. W. Dutton The Rate of the CD8-Dependent Initial Reduction in Tumor Volume Is Not Limited by Contact-Dependent Perforin, Fas Ligand, or TNF-Mediated Cytolysis J. Immunol., August 1, 2004; 173(3): 1738 - 1743. [Abstract] [Full Text] [PDF]
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 J. A. DeVoti, B. M. Steinberg, D. W. Rosenthal, L. Hatam, A. Vambutas, A. L. Abramson, M. J. Shikowitz, and V. R. Bonagura Failure of Gamma Interferon but Not Interleukin-10 Expression in Response to Human Papillomavirus Type 11 E6 Protein in Respiratory Papillomatosis Clin. Vaccine Immunol., May 1, 2004; 11(3): 538 - 547. [Abstract] [Full Text] [PDF]
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L. A. O'Mara and P. M. Allen Pulmonary Tumors Inefficiently Prime Tumor-Specific T Cells J. Immunol., January 1, 2004; 172(1): 310 - 317. [Abstract] [Full Text] [PDF]
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 E. A. Holly, C. A. Eberle, and P. M. Bracci Prior History of Allergies and Pancreatic Cancer in the San Francisco Bay Area Am. J. Epidemiol., September 1, 2003; 158(5): 432 - 441. [Abstract] [Full Text] [PDF]
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T. Woodberry, J. Gardner, S. L. Elliott, S. Leyrer, D. M. Purdie, P. Chaplin, and A. Suhrbier Prime Boost Vaccination Strategies: CD8 T Cell Numbers, Protection, and Th1 Bias J. Immunol., March 1, 2003; 170(5): 2599 - 2604. [Abstract] [Full Text] [PDF]
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M. P. Rubinstein, A. N. Kadima, M. L. Salem, C. L. Nguyen, W. E. Gillanders, M. I. Nishimura, and D. J. Cole Transfer of TCR Genes into Mature T Cells Is Accompanied by the Maintenance of Parental T Cell Avidity J. Immunol., February 1, 2003; 170(3): 1209 - 1217. [Abstract] [Full Text] [PDF]
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S. Schwaiger, A. M. Wolf, P. Robatscher, B. Jenewein, and B. Grubeck-Loebenstein IL-4-Producing CD8+ T Cells with a CD62L++(bright) Phenotype Accumulate in a Subgroup of Older Adults and Are Associated with the Maintenance of Intact Humoral Immunity in Old Age J. Immunol., January 1, 2003; 170(1): 613 - 619. [Abstract] [Full Text] [PDF]
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 K.-J. Malmberg, R. Lenkei, M. Petersson, T. Ohlum, F. Ichihara, B. Glimelius, J.-E. Frodin, G. Masucci, and R. Kiessling A Short-Term Dietary Supplementation of High Doses of Vitamin E Increases T Helper 1 Cytokine Production in Patients with Advanced Colorectal Cancer Clin. Cancer Res., June 1, 2002; 8(6): 1772 - 1778. [Abstract] [Full Text] [PDF]
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) or LCMV33–41-loaded DC (•) or were left
uninjected (
). After 7 days, mice were challenged with 5 x
105 LL-LCMV tumor cells injected s.c. in the opposite
flank. Tumor size is expressed as the mean product of bisecting tumor
diameters ± SEM. Three to five mice were used in each
experimental group. The experiment was repeated at least three times
with similar results. Data from one representative experiment are
shown.
,
EL-4 targets plus LCMV33–41; no symbol, EL-4 target cells
left unpulsed. The result from only one of the three unpulsed controls
is shown because all three gave overlapping profiles. The experiment
was repeated at least four times with similar results. Data from one
representative experiment are shown.
, Cytokines
produced by Tc1 cells;