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,
Departments of
*
Obstetrics and Gynecology and
Pathology, National Taiwan University Hospital, and
Graduate Institute of Immunology, National Taiwan University College of Medicine, Taipei, Taiwan
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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in cancer-encountered T cells. The regulatory effects of
cervical cancer cells are mediated mainly by IL-10, and TGF-
plays
only a synergistic role. The cancer-derived effects can be reversed by
neutralizing anti-IL-10 and anti-TGF- Abs. IL-10 and TGF-
are present in cancer tissue and weakly expressed in precancerous
tissue, but not in normal cervical epithelial cells. Our study strongly
suggests important regulatory roles of IL-10 and TGF- in
cancer-mediated immunosuppression. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, or a Tc2 subset that produces
IL-4, IL-5, and IL-10 (5, 6, 7, 8, 9). In human T cell clones,
Th1/Tc1 cells preferentially develop during infections with
intracellular bacteria and enhance phagocyte-mediated immunity, whereas
Th2/Tc2 cells predominate during metazoan parasite infestations and in
atopic diseases (4, 5, 6). Although both types of
CD8+ T cell subsets are thought to possess a
cytolytic function in vitro, there is substantial evidence indicating
that CD8+ T cells secreting type 1 and type 2
cytokines in vivo might have great relevance to immune responses
(10, 11, 12, 13). In a recent study of TCR-transgenic mice, Tc2
has been demonstrated to be less protective than Tc1 against pulmonary
influenza virus infection (13). Because Tc cells play a
central role in cellular immunity against cancers, alterations of
Tc1/Tc2 subsets in the tumor microenvironment may change host
anti-tumor immune responses (8, 9, 10, 11, 12). In human cervical cancer (CC) as well as in other cancers, depressed anti-tumor immunity of Tc cells has been observed (14, 15, 16). The cytotoxic potential of freshly isolated tumor-infiltrating lymphocytes (TILs) is usually not expressed (16, 17, 18). This suggests the possible existence of as yet undefined mechanisms that affect the anti-tumor capacity of these TILs in the tumor milieu. Previously, we have demonstrated that cancer cells may alter the functional composition of anti-tumor effector cells (e.g., CD3+CD25+ T cells) within the tumor milieu (19, 20). We have further illustrated that a decreased proportion of CD4+ T cells is responsible for the reversed CD4/CD8 ratio of TILs in human CC (20). Because the tumor-reactive CD4+ T cells can be activated through the MHC class II pathway to produce lymphokines and amplify the cytotoxic activity of CD8+ T cells (21, 22), a decreased proportion of CD4+ T cells with a reversed CD4/CD8 ratio in vivo may result from functional alterations of T cells within the tumor microenvironment.
In this study, we examined the functional characteristics of TILs by
investigating their cytokine expression (23, 24). We used
an in vitro mixed autologous lymphocyte and tumor-cell coculture (MLTC)
model to evaluate the kinetic expression of cytokines by
cancer-encountered T cells (25). Our results demonstrate
for the first time that activated T cells in the human cancer milieu
predominantly express a Th2/Tc2 phenotype, and that cancer cells can
directly drive the tumor-encountered T cells toward Th2/Tc2 polarity
through an IL-10- and TGF--mediated pathway.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Patients with stage Ib CC admitted for operation were prospectively enrolled in this study. A complete history was obtained for each patient. Informed consent was obtained for collecting the materials in this study. After radical hysterectomy, the surgical specimens were carefully examined by experienced pathologists. Tissue specimens were excised aseptically immediately after operation from at least four different tumor sites and two sites of normal cervical epithelium of the same tumor patients. Fragments of tissue were washed with PBS for removal of contaminated blood. CC cells, cervical epithelial cells, and TILs were isolated according to methods described previously (19, 20, 25). For matched comparison, venous blood of each patient was obtained before operation and transferred to test tubes containing heparin. PBMCs were isolated by Ficoll-Hypaque (1.077 density) and resuspended at a concentration of 1 x 106 cells/ml in RPMI 1640 medium. The recovered cells were checked and counted for viability with the trypan blue staining method. Purified TILs and PBMCs (1 x 106 cells/ml) were activated in medium containing PMA at 25 ng/ml, ionomycin at 1 µg/ml, and brefeldin A at 10 µg/ml. The activated cells were incubated in a 37°C, 5% CO2 humidified incubator for 18 h. The activation status was monitored by surface staining of CD69.
Surface and intracellular staining
Monoclonal Abs labeled with FITC, PE, and PerCP were used for
triple-color flow cytometry as previously described (19, 20, 25). For surface Ag staining, 20 µl anti-CD4-FITC,
anti-CD8-FITC, and/or anti-CD3-PerCP (BD Immunocytometry
Systems, San Jose, CA) were used for every set of 1 x
106 activated T cells. After incubation for 30
min, cells were washed twice with 1 ml lysing buffer, and fixed with 1
ml fixation buffer containing 1% paraformaldehyde. Cells were washed
twice with 500 µl permeabilization buffer containing 0.5% saponin,
and stained with 20 µl fluorescence-conjugated anti-cytokine mAbs
for an additional 30 min at room temperature. Anti-IFN--PE,
anti-IL-5-PE (BD PharMingen; San Diego, CA), and anti-IL-4-FITC
plus anti-IFN--PE (BD Immunocytometry Systems) were used for
intracellular staining. Anti-mouse cytosol-IgG1-FITC + cytosol-IgG2a-PE
was used as negative control. Triple-color flow cytometry was performed
on a FACSCalibur (BD Biosciences, Mountain View, CA). Data were
acquired with CellQuest software (BD Biosciences) by the use of
forward-scatter/side-scatter thresholds with 10,000 gated events. We
used Leukogate to measure the proportion of lymphocytes in the
sample being studied. In triple-color assays with cytokine expression,
data were further gated on FL3+ cells with a CD3
trigger for evaluation of T cell subsets.
Purification of CD8+ cytotoxic T lymphocytes
CD8+ T cells were isolated from TILs by use of an indirect magnetic labeling system with a MACS (Miltenyi Biotec, Gladbach, Germany). The cells were suspended in PBS medium containing 20 µl of Hepten-Ab Cocktail (containing CD4, CD11b, CD16, CD19, CD36, and CD56 Abs; Miltenyi Biotec) and incubated in a cold room (6-12°C) for 10 min. After repeated washing, 20 µl of MACS anti-hepten microbeads per 107 total cells was added. The suspension was further incubated for 15 min at 6-12°C, then washed carefully by the addition of 10–20x the labeling volume of the washing buffer. The magnetically labeled cells were passed through a MACS separator in a magnetic field. The column was rinsed with an adequate volume of buffer and repeatedly eluted. The effluent was collected as negative fraction, representing the enriched CD8+ T cell fraction. The cell purity was checked by flow cytometry after labeling with anti-CD8-FITC.
RT-PCR analysis of mRNA expression
We measured the cytokine mRNA expression in purified
CD8+ T cells derived from TILs and PBMCs, CC
cells, and normal cervical epithelial cells by RT-PCR. The mRNAs of
cytokines involved in the functional T cell development were examined.
Total cellular RNA was extracted and reverse-transcribed (0.1–0.5 mg
RNA) in the presence of IL-4, IL-10, IL-12, IFN-, TNF-
, and
TGF- oligonucleotide primers (CLONTECH Laboratories, Palo Alto, CA).
Human -actin primers were used as positive controls. The PCR was
performed in a DNA thermal cycler (GeneAmp DNA thermal cycler 480;
PerkinElmer, Norwalk, CT) for 36 cycles of denaturation at 94°C for
40 s, annealing at 55°C for 1 min, followed by polymerization at
72°C for 1 min. PCR-amplified products were separated on an agarose
gel and visualized by staining with ethidium bromide.
Mixed lymphocyte and tumor cell coculture assay
To address the possible role of cancer cells in T cell polarity,
we cultured PBMCs with autologous cancer cells or normal cervical
epithelial cells (25). The procedures outlined were
performed in duplicate in 12 mm x 75 mm capped polystyrene test
tubes. PBMCs were activated with 10 µg/ml of PHA or anti-CD3 (1
µg/ml) plus anti-CD28 (1 µg/ml) and goat anti-mouse
cross-linkers (2 µg/ml; Immunotech, Marseille, France) in a 37°C,
5% CO2 humidified incubator. A constant number
(5 x 105) of PBMCs were cultured with
autologous cancer cells (1 x 106) in the
MLTC group. In the control group of mixed autologous lymphocytes and
noncancerous epithelial cells coculture (MLNC), 5 x
105 activated T cells were cultured with 1
x 106 autologous normal cervical epithelial
cells. The supernatant on days 1, 3, and 5 of coculture was collected
at the same intervals and checked for the presence of IL-4 and IFN-
by a standard ELISA. The cytokine expressions of T cells in MLTC and
MLNC were measured by an intracellular staining method and flow
cytometry as described in the previous sections.
ELISA for IL-4 and IFN- measurement
Supernatants of MLTC and MLNC were collected in small aliquots.
The levels of IL-4 and IFN- were measured with a commercial ELISA
kit (Endogen, Woburn, MA). A spectrophotometer set to 450–550 nm was
used for measuring the OD of each well of the 96-well polystyrene
microtiter plate. Triplicate readings were averaged and expressed in
units of picograms per milliliter.
Expression of IL-10 and TGF- in CC tissue
An avidin-biotin-peroxidase complex immunohistochemical staining
method was performed for examination of the expression pattern of IL-10
and TGF- in formalin-fixed and paraffin-embedded tissue sections.
The anti-IL-10 Ab was obtained from Santa Cruz Biotechnology (Santa
Cruz, CA). The anti-TGF- Ab was obtained from Serotec
(Kidlington, Oxford, U.K.). The specificities of both Abs were provided
by the manufacturers.
Effects of anti-IL-10 and anti-TGF- Abs on T cell
polarity
To clarify the functional role of cancer-derived IL-10 and
TGF- on T cell polarity, we added anti-IL-10 (BD PharMingen) and
anti-TGF- Abs (Serotec) to the previously established MLTC. Cancer
cells (1 x 106) were first cultured in 1 ml
of culture medium containing 10 µg/ml of anti-IL-10 and/or
anti-TGF- Abs for 2 h and then cocultured with activated T
cells (5 x 105 cells) in the T cell and
cancer cell coculture (T+C) group. For matched comparison, a
constant number (5 x 105) of activated T
cells was cultured as the T-only group. Mouse anti-human IgG1 and
rat anti-human IgG (10 µg/ml) were used as isotype Ab controls.
The supernatant of the coculture was collected at the same intervals
and checked for the presence of IL-4 and IFN-.
Direct effects of recombinant IL-10 and TGF- on T cell polarity
We further directly explored the functional role of IL-10 and
TGF- on T cell polarity. CD3+ T cells were
purified by MACS and activated with anti-CD3 (1 µg/ml) plus
anti-CD28 (1 µg/ml) and goat anti-mouse cross-linkers (2
µg/ml) (Immunotech) as previously mentioned. The activated cells were
labeled with anti-CD69-PE and checked by flow cytometry.
Recombinant IL-10 and TGF- (PeproTech, Rocky Hill, NJ) were added to
the cultures of activated T cells (5 x 105)
at a final concentration of 5 and 1 ng/ml, respectively. For matched
comparison, a constant number (5 x 105) of
activated T cells was cultured as the T-only group. In addition to the
previously established MLTC (T+C group), anti-IL-10 and
anti-TGF- Abs were added in another set of MLTC (T+C plus
anti-IL-10/anti-TGF- group). The cytokine expression of
gated CD3+ T cells was measured on day 3 by the
intracellular staining method and flow cytometry as described in the
previous sections. The supernatant of the coculture was collected at
the same time and checked for the presence of IL-4 and IFN-. Studies
were performed in triplicate.
Statistical analysis of data
Data analysis was performed with a statistical analysis system (R.6.12; SAS Institute, Cary, NC). The Wilcoxon signed-ranks test was used for comparison of the median expression ratio of cytokine expression in CD3+, CD3+CD4+, and CD3+CD8+ T cell subsets between TILs and PBMCs. Kinetic data of MLTC and MLNC were analyzed by a generalized estimation equation for correlated data of repeated measurements and were expressed as mean ± SD. Statistical significance was defined by a p value <0.05.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Histologic examination of the specimens revealed that all CC cases
were squamous cell carcinomas. The viability of immunocytes was
90–95% at the completion of the isolation procedure in all cases.
In triple-color flow cytometric assays with dual-cytokine intracellular
staining of eight CC cases, gated CD3+ T cells
(Fig. 1
, a and b)
derived from TILs expressed mainly IL-4, but not IFN- (Fig. 1c). In contrast, prominent expression of IFN-, but not
IL-4, was observed in activated CD3+ PBMCs (Fig. 1d). Subsequent staining of dual-surface phenotyping in
combination with cytokine intracellular staining showed that the
expression of these cytokines was restricted to particular subsets of T
cells (Fig. 1, e–h). Prominent expression of IL-5 with low
expression of IFN- was demonstrated in both gated
CD3+CD4+ (data not shown)
and CD3+CD8+ TIL subsets
(Fig. 1, e and f) derived from CC tissue.
Contrarily, prominent expression of IFN- with low expression of IL-5
was detected in both activated
CD3+CD4+ (data not shown)
and CD3+CD8+ PBMCs (Fig. 1, g and h). Flow cytometric analyses further
defined the ratios of intracellular cytokine expression in individual T
cell subsets (Fig. 2, a and
b). The median expression ratio of IL-5 on
CD3+CD4+ cells was
significantly higher in TILs than in PBMCs (60.71 vs 0.25%,
n = 8, p < 0.01). A similar finding
was obtained with CD3+CD8+
cells (66.73 vs 0.28%, n = 8, p <
0.01), which signified the predominant Th2/Tc2 patterns of cytokine
expression in TILs.
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mRNA expression in
CD8+ T cells isolated from TILs and activated
PBMCs (Fig. 2
c). Similar to the result obtained with
intracellular staining, we observed the abundant expression of IL-4
mRNA, but low expression of IFN- mRNA, in CD8+
TILs. The PBMC-derived CD8+ T cells prominently
expressed IFN- mRNA and low expression of IL-4 mRNA. The results
emphasized that tumor-encountered T cells in the cancer milieu have a
predominant Th2/Tc2 phenotype of mRNA expression and cytokine
production. Determination of the effects of cancer cells on TILs
It is possible that CC cells can direct the differentiation of
activated T cells toward Th2/Tc2 polarity. To clarify this possibility,
we used a previously established in vitro cancer-host interaction model
and analyzed the cytokine production of IL-4 and IFN- in the
supernatants of MLTC and MLNC (25). In six sets of
repeated experiments, the mean concentration of IFN- in MLNC
supernatants was 201.2 ± 16.4 pg/ml and increased >1- and 2-fold
by days 3 and 5, respectively. However, the mean concentration of
IFN- in the supernatants of MLTC was significantly lower than that
of MLNC in day 5 cocultures (n = 6, p
< 0.01) (Fig. 3a). The mean
concentration of IL-4 in MLNC supernatants was 64.1 ± 6.6 pg/ml
and increased by <1-fold by days 3 and 5. However, the mean
concentration of IL-4 in the supernatants of MLTC was significantly
higher than that in MLNC in day 3 and day 5 cocultures (increased >3-
and 4-fold, respectively) (n = 6, p <
0.001) (Fig. 3b). It became evident that CC cells could
promote the production of IL-4 and possibly abrogate the production of
IFN- in cancer-encountered T cells.
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To examine the possible cancer-derived immunosuppressive factors
or cytokines that regulate the functional development or polarization
of cancer-encountered T cells, we further compared the mRNA expression
of immunoregulatory cytokines, including IL-4, IL-10, IL-12, IFN-,
TNF-, and TGF-, in 30 samples of CC cells. IL-4, IL-12, IFN-,
and TNF- mRNAs were not detected in either cancer or epithelial
cells. However, TGF- and IL-10 mRNAs were highly expressed in
the CC cells, but not in the normal cervical epithelial cells (Fig. 3c). We then studied the existence of TGF- and
IL-10 in CC by immunohistochemical staining of CC tissues from 47
patients. TGF- and IL-10 were abundantly expressed in most, if not
all cancer cells, but were not or only very weakly expressed in normal
cervical epithelial or stromal cells (Fig. 4, a and b). The
expression of TGF- and IL-10 became obvious in transitional sections
from normal cervical epithelium to carcinoma-in-situ and invasive
cancer areas (Fig. 4, c and d). Unlike the
expression of TGF- and IL-10 in cancer cells, the expression of
other types of immunoregulatory cytokines was not detected.
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in T cell
polarity
To characterize the possible role of IL-10 and TGF- in the
polarity of cancer-encountered T cells further, we directly incubated
anti-IL-10 and anti-TGF- Abs in the autologous MLTC. Using
the kinetic experiments on MLTC described above, we determined whether
specific blocking Abs could reverse or limit the cancer-induced
effects. PBMCs were fully activated as matched controls (activated
T-cells-only group). In six sets of matched experiments after 5 days of
coculture, the mean concentration of IFN- in the supernatants of
MLTC was significantly lower than that in the T-cells-only group
(287.05 ± 17.12 vs 811.40 ± 30.96 pg/ml, n
= 6, p < 0.01) (Fig. 5a). The mean
concentration of IL-4 in the MLTC group was 3-fold that in the
activated T-cells-only group (347.06 ± 13.61 vs 103.39 ±
18.34 pg/ml, n = 6, p < 0.01) (Fig. 5b).
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in the supernatants of MLTC was observed in day 3 and day 5
cocultures (Fig. 5a). The mean concentration of IL-4 in the
supernatants of MLTC was synchronously reduced on day 3 and day 5
cocultures (48 and 57% decrease, respectively) (Fig. 5b).
The effect of anti-TGF- Ab on cancer-induced IL-4 production was
not as obvious as that of the anti-IL-10 Ab, with only 19 and 25%
decreases in day 3 and day 5 cocultures, respectively. However, adding
both anti-IL-10 and anti-TGF- Abs in MLTC achieved a maximal
reduction of IL-4 in the supernatants, being 65 and 70% reductions in
day 3 and day 5 cocultures, which were close to the mean concentrations
of IL-4 in the supernatants of the T-cells-only group (Fig. 5b). The effects of anti-IL-10 and anti-TGF- Abs
were also observed in the patterns of IFN- in the supernatants of
MLTC (Fig. 5a). The isotype controls did not alter the
production of IL-4 or IFN- in the MLTC (data not shown). Our data
illustrated that cancer cells promote the production of IL-4 and
abrogate the production of IFN- in cancer-encountered T cells. These
cancer-derived effects can be reversed by anti-IL-10 and
anti-TGF- Abs.
Compatible cancer-derived effects and recombinant IL-10/TGF- on
T cell polarity
To clarify the possible synergistic effect of cancer-derived IL-10
and TGF- on the polarity of T cells, we further incubated activated
T cells with recombinant IL-10 and TGF- in addition to the
established autologous MLTC. Using the kinetic experiments on MLTC and
the intracellular staining technique described above, we directly
determined the cancer-derived effects on cancer-encountered T cells. In
eight sets of matched experiments after 3 days of coculture, the median
expression ratio of IL-4 was significantly higher in the MLTC (T+C
group) than in the activated T-cells-only group (77.28 vs 0.31%,
n = 8, p < 0.01). The median
expression ratio of IFN- in MLTC was lower than that in the
T-cells-only group (0.46 vs 87.57%, n = 8,
p < 0.01). The median expression ratio of IL-4 in T
cells treated with recombinant IL-10 and TGF- was consistent with
that in MLTC (72.53 vs 77.28%). Adding both anti-IL-10 and
anti-TGF- Abs in MLTC significantly reduced the expression of
IL-4 in CD3+ T cells (5.8 vs 77.28%,
n = 8, p < 0.01). Similar results were
seen in the measurements of IL-4 and IFN- of the coculture
supernatants. The mean concentration of IL-4 in supernatants of the
MLTC group was similar to that of culture of activated T cells in
recombinant IL-10 and TGF- (202.78 ± 74.51 and 199.29 ±
79.77 pg/ml, respectively). Anti-IL-10 and anti-TGF- Abs in MLTC
decreased the mean concentration of IL-4 to the levels of the
T-cells-only group, being 56.97 ± 11.94 and 58.49 ± 8.78
pg/ml, respectively. It became evident that human cancer cells have
well-matched effects as that of recombinant IL-10 and TGF- on
driving the polarity of cancer-encountered T cells. Importantly, adding
anti-IL-10 and anti-TGF- Abs can reverse the cancer-derived
effects.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In the present study, we directly demonstrated that the T cell (both
CD4+ and CD8+ cell)
polarity can be driven toward the Th2/Tc2 subset by IL-10 and TGF-
from human cancer cells. In triple-color flow cytometric assays with
dual-cytokine intracellular staining, CD3+ T
cells derived from TILs expressed mainly type 2 cytokine patterns. We
further defined the ratios of intracellular cytokine expression in
individual CD3+CD4+ and
CD3+CD8+ T cell subsets in
TILs and signified the predominant Th2/Tc2 patterns of cytokine
expression in the cancer milieu. Using an in vitro coculture model of
MLTC, we directly demonstrated that human cancer cells could promote
the production of IL-4 and possibly abrogate the production of IFN-
in cancer-encountered T cells through the synergistic effect of IL-10
and TGF-.
Our findings link both in vivo data that, for the first time, reveal polarity in TILs and show in vitro for the first time that human cancer cells can drive encountered T cells toward Tc2 polarity by cancer-derived mediators. Polarization toward Th2/Tc2 subsets may signify a down-regulation of the cytotoxic potential of TILs and/or render the tumor cells resistant to TIL-mediated cytolysis. Our results confirm that cancer cells may attain immune escape by changing the effector composition of the host immune cells. We further explore the issues of proposed cancer-host immune interactions based on previous in vivo murine models and in vitro cell line results (27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40).
TGF- and IL-10 are secreted spontaneously by a variety of human
malignant tumors, including breast, colon, ovary, cutaneous
basal/squamous cancers, metastatic melanoma, fibrosarcoma, and
pancreatic carcinoma (27, 28, 29, 30, 31, 32, 33, 34, 35). The mechanisms responsible
for the increased expression by tumor cells are not known. It has been
suggested that tumor-derived IL-10 may be associated with the
down-regulation of anti-tumor immunity by inhibiting the
cytotoxicity of CTLs, tumor Ag presentation, and Th1 cell development
(27, 28, 35, 36). However, related studies in the
literature showed indirect evidences of either elevated IL-4 and IL-10
mRNA, or mixed type 2 cytokine patterns in cancers (28, 37, 38). In the present study, we directly demonstrated predominant
in vivo Th2/Tc2 patterns of cytokine expression in subsets of TILs, and
stratified that tumor-derived IL-10 promotes a shift toward Th2/Tc2
polarity. The immunosuppressive role of IL-10 has been emphasized as a
strategy used by tumors to escape immunosurveillance (39).
A recent study by Steinbrink et al. (40), showing that
IL-10-treated human dendritic cells can induce anergy in tumor-specific
cytotoxic CD8+ T cells and result in failure to
lyse tumor cells, further elucidates the down-regulatory role of IL-10
in tumor-mediated immunosuppression.
TGF- has pleiotropic and potent immunosuppressive effects in the
regulation of various physiologic processes. Tumor-derived TGF- has
been shown to down-regulate the generation of tumor-specific cytotoxic
lymphocytes (41, 42, 43) and the secretion of immunoregulatory
cytokines by T cells (42). TGF- can be a strong
costimulator that promotes a shift toward Th2 polarity in rat activated
T cells (44). In a tumor-bearing murine model, TGF- has
been shown to shift the Th1/Th2 balance through a direct,
IL-10-mediated pathway (45). By secreting IL-10 and
TGF-, tumor cells can affect the activation, proliferation, and
differentiation of TILs in a synergistic manner. We have demonstrated
in the MLTC model that human cancer cells can drive T cell polarity
toward the Th2/Tc2 subsets by the synergistic effects of TGF- and
IL-10. The cancer-derived IL-10 and TGF- may play a synergistic role
in the induction of immune privilege in local cancer milieu
(46). It is worthy of note that both IL-10 and TGF- had
expressions that progressed from carcinoma in situ to invasive cancer,
but were not expressed in normal cervical epithelium of transitional
sections (Fig. 4, a–d). The correlated presence of
IL-10/TGF- and Th2/Tc2 polarity of TILs in cancer tissue
demonstrates that cancer-derived mediators can be active in modulating
T cell functions.
In conclusion, we demonstrated that activated T cells in a human CC
milieu predominantly expressed the Th2/Tc2 phenotype. We showed that
cancer cells could drive the tumor-encountered T cells toward Th2/Tc2
polarity, which may be attributed to the prominent IL-10 and TGF-
expression in CC cells.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Su-Cheng Huang, Department of Obstetrics and Gynecology, National Taiwan University Hospital, No. 7, Chung-Shan South Road, Taipei 100, Taiwan. E-mail address: bcsheu{at}ha.mc.ntu.edu.tw
3 Abbreviations used in this paper: Tc, T cytotoxic (cell); TIL, tumor-infiltrating lymphocyte; CC, cervical cancer; MLTC, mixed autologous lymphocyte and tumor-cell coculture; MLNC, mixed autologous lymphocytes and noncancerous epithelial cells coculture; T+C, T cell and cancer cell coculture.
Received for publication March 16, 2001. Accepted for publication June 18, 2001.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
and in interleukin-4-expressing cells. Eur. J. Immunol. 24:1097.[Medline]
1. Endocrinology 130:2476.[Abstract]
expression inhibits cell proliferation in vitro, yet increases tumorigenicity and tumor growth of Meth A carcinoma cells. Cancer Res. 53:4391. as prognostic factors in breast carcinoma. Int. J. Cancer 74:492.[Medline]
and interleukin-10 contribute a systemic Th2 immune phenotype in pancreatic carcinoma patients. Am. J. Pathol. 155:537. on IL-2- and IL-4-dependent T cell-cycle progression. J. Immunol. 144:1767.[Abstract]
on the immune system: implications for cancer immunotherapy. Leukemia 13:1188.[Medline]
1 cDNA escapes immune surveillance. Proc. Natl. Acad. Sci. USA 87:1486.1, a strong costimulator of rat T-cell activation promoting a shift towards a Th2-like cytokine profile. Immunol. Lett. 67:131.
contributes to the shift toward Th2-type responses through direct and IL-10-mediated pathways in tumor-bearing mice. J. Immunol. 156:73.[Abstract]
and IL-10 in the induction of immune privilege. J. Immunol. 160:2089.This article has been cited by other articles:
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B.-C. Sheu, S.-H. Chiou, H.-H. Lin, S.-N. Chow, S.-C. Huang, H.-N. Ho, and S.-M. Hsu Up-regulation of Inhibitory Natural Killer Receptors CD94/NKG2A with Suppressed Intracellular Perforin Expression of Tumor-Infiltrating CD8+ T Lymphocytes in Human Cervical Carcinoma Cancer Res., April 1, 2005; 65(7): 2921 - 2929. [Abstract] [Full Text] [PDF]
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A. de Jong, M. I. E. van Poelgeest, J. M. van der Hulst, J. W. Drijfhout, G. J. Fleuren, C. J. M. Melief, G. Kenter, R. Offringa, and S. H. van der Burg Human Papillomavirus Type 16-Positive Cervical Cancer Is Associated with Impaired CD4+ T-Cell Immunity against Early Antigens E2 and E6 Cancer Res., August 1, 2004; 64(15): 5449 - 5455. [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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M. J. Dobrzanski, J. B. Reome, J. A. Hollenbaugh, and R. W. Dutton Tc1 and Tc2 Effector Cell Therapy Elicit Long-Term Tumor Immunity by Contrasting Mechanisms That Result in Complementary Endogenous Type 1 Antitumor Responses J. Immunol., February 1, 2004; 172(3): 1380 - 1390. [Abstract] [Full Text] [PDF]
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 K.-H. Chao, M.-Y. Wu, J.-H. Yang, S.-U. Chen, Y.-S. Yang, and H.-N. Ho Expression of the interleukin-2 receptor {alpha} (CD25) is selectively decreased on decidual CD4+ and CD8+ T lymphocytes in normal pregnancies Mol. Hum. Reprod., July 1, 2002; 8(7): 667 - 673. [Abstract] [Full Text] [PDF]
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) were analyzed by ELISA. a, The mean concentration
of IFN-
), anti-IL-10 (
), and
anti-IL-10 + anti-TGF-
). Activated T cells (T
cell-only) were used as controls (