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Non-redundant role for IL-12 and IL-27 in modulating Th2 polarization of carcinoembryonic antigen specific CD4 T cells from pancreatic cancer patients.

Tassi E, Braga M, Longhi R, Gavazzi F, Parmiani G, Di Carlo V, Protti MP - PLoS ONE (2009)

Bottom Line: This has negative effects in both effective anti-tumor immunity and further favoring fibrinogenesis.We found that only the combination of IL-12 and IL-27 modified the polarization of Th2 effectors by both reduction of IL-5, GM-CSF and IL-13 and induction of IFN-gamma production, which lasted after cytokine removal.In agreement with the data obtained with the clones, we found that the combined treatment functionally modulated the Th2 polarization of CEA-specific CD4(+) T cells and enhanced pre-existing Th1 type immunity.

View Article: PubMed Central - PubMed

Affiliation: Tumor Immunology Unit, San Raffaele Scientific Institute, Milan, Italy.

ABSTRACT

Background: Pancreatic cancer is a very aggressive disease with dismal prognosis; peculiar is the tumor microenvironment characterized by an extensive fibrotic stroma, which favors rapid tumor progression. We previously reported that pancreatic cancer patients have a selective Th2 skew in the anti-carcinoembryonic antigen (CEA) CD4(+) T cell immunity, which correlates with the presence of a predominant GATA-3(+) tumor lymphoid infiltrate. This has negative effects in both effective anti-tumor immunity and further favoring fibrinogenesis. Aim of this study was to evaluate whether the Th2 polarization of CEA-specific CD4(+) T cells from pancreatic cancer patients is stable or can be reverted by immunomodulating cytokines.

Methodology/principal findings: We first evaluated the influence of IL-12 and IL-27, as single agents and in association, on the polarization of CEA-specific Th2 CD4(+) T cell clones from a pancreatic cancer patient. We found that only the combination of IL-12 and IL-27 modified the polarization of Th2 effectors by both reduction of IL-5, GM-CSF and IL-13 and induction of IFN-gamma production, which lasted after cytokine removal. Second, we evaluated the effect of the combined treatment on polyclonal CEA-specific CD4(+) T cells in short-time re-stimulation assays. In agreement with the data obtained with the clones, we found that the combined treatment functionally modulated the Th2 polarization of CEA-specific CD4(+) T cells and enhanced pre-existing Th1 type immunity.

Conclusions/significance: Collectively, our results demonstrate that tumor antigen specific Th2 CD4(+) T cells in pancreatic cancer are endowed with functional plasticity. Hence, loco-regional cytokines delivery or targeted therapy based on antibodies or molecules directed to the tumor stroma might improve anti-tumor immunity and ameliorate fibrosis, without systemic toxicity.

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Modulation of Th2 polarization of CEA177–189/355–367 specific CD4+ T cells by combined IL-12 and IL-27 treatment.CD4+ T cells were cultured with the relevant peptide in the absence (basal) or in the presence of IL-12 (5 ng/ml), as single agent; or IL-27 (100 ng/ml), as single agent; or combined IL-12 and IL-27 in a 2-day stimulation assay and then tested for cytokine release by CBA (IL-5 and IFN-γ) or ELISA (IL-13 and GM-CSF). The data for IL-13 and GM-CSF are means of duplicate determination±SD. The basal level of cytokines secretion of CD4+ T cells in the presence of LCL only was subtracted from the sample values and was comprised between 0 and 0,006 ng/ml. The data are representative of five experiments.
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pone-0007234-g003: Modulation of Th2 polarization of CEA177–189/355–367 specific CD4+ T cells by combined IL-12 and IL-27 treatment.CD4+ T cells were cultured with the relevant peptide in the absence (basal) or in the presence of IL-12 (5 ng/ml), as single agent; or IL-27 (100 ng/ml), as single agent; or combined IL-12 and IL-27 in a 2-day stimulation assay and then tested for cytokine release by CBA (IL-5 and IFN-γ) or ELISA (IL-13 and GM-CSF). The data for IL-13 and GM-CSF are means of duplicate determination±SD. The basal level of cytokines secretion of CD4+ T cells in the presence of LCL only was subtracted from the sample values and was comprised between 0 and 0,006 ng/ml. The data are representative of five experiments.

Mentions: Second, we evaluated the effect of the combined treatment with the two cytokines (Fig. 3). When used at 5 ng/ml, IL-12, as a single agent, induced 61% inhibition of IL-5 and had no effect on IL-13 and GM-CSF, while IFN-γ production had almost a 10-fold increase. IL-27 treatment alone at the highest concentration showed 78%, 30% and 53% inhibition of IL-5, IL-13 and GM-CSF, respectively; while only 1,4-fold increase in IFN-γ production. When used in combination at the best concentrations, a synergistic effect was observed in the inhibition of IL-5 (91%) and IL-13 (48%), while, as expected from the results of the single agents, GM-CSF secretion was not further inhibited and IFN-γ production was not further increased.


Non-redundant role for IL-12 and IL-27 in modulating Th2 polarization of carcinoembryonic antigen specific CD4 T cells from pancreatic cancer patients.

Tassi E, Braga M, Longhi R, Gavazzi F, Parmiani G, Di Carlo V, Protti MP - PLoS ONE (2009)

Modulation of Th2 polarization of CEA177–189/355–367 specific CD4+ T cells by combined IL-12 and IL-27 treatment.CD4+ T cells were cultured with the relevant peptide in the absence (basal) or in the presence of IL-12 (5 ng/ml), as single agent; or IL-27 (100 ng/ml), as single agent; or combined IL-12 and IL-27 in a 2-day stimulation assay and then tested for cytokine release by CBA (IL-5 and IFN-γ) or ELISA (IL-13 and GM-CSF). The data for IL-13 and GM-CSF are means of duplicate determination±SD. The basal level of cytokines secretion of CD4+ T cells in the presence of LCL only was subtracted from the sample values and was comprised between 0 and 0,006 ng/ml. The data are representative of five experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2749205&req=5

pone-0007234-g003: Modulation of Th2 polarization of CEA177–189/355–367 specific CD4+ T cells by combined IL-12 and IL-27 treatment.CD4+ T cells were cultured with the relevant peptide in the absence (basal) or in the presence of IL-12 (5 ng/ml), as single agent; or IL-27 (100 ng/ml), as single agent; or combined IL-12 and IL-27 in a 2-day stimulation assay and then tested for cytokine release by CBA (IL-5 and IFN-γ) or ELISA (IL-13 and GM-CSF). The data for IL-13 and GM-CSF are means of duplicate determination±SD. The basal level of cytokines secretion of CD4+ T cells in the presence of LCL only was subtracted from the sample values and was comprised between 0 and 0,006 ng/ml. The data are representative of five experiments.
Mentions: Second, we evaluated the effect of the combined treatment with the two cytokines (Fig. 3). When used at 5 ng/ml, IL-12, as a single agent, induced 61% inhibition of IL-5 and had no effect on IL-13 and GM-CSF, while IFN-γ production had almost a 10-fold increase. IL-27 treatment alone at the highest concentration showed 78%, 30% and 53% inhibition of IL-5, IL-13 and GM-CSF, respectively; while only 1,4-fold increase in IFN-γ production. When used in combination at the best concentrations, a synergistic effect was observed in the inhibition of IL-5 (91%) and IL-13 (48%), while, as expected from the results of the single agents, GM-CSF secretion was not further inhibited and IFN-γ production was not further increased.

Bottom Line: This has negative effects in both effective anti-tumor immunity and further favoring fibrinogenesis.We found that only the combination of IL-12 and IL-27 modified the polarization of Th2 effectors by both reduction of IL-5, GM-CSF and IL-13 and induction of IFN-gamma production, which lasted after cytokine removal.In agreement with the data obtained with the clones, we found that the combined treatment functionally modulated the Th2 polarization of CEA-specific CD4(+) T cells and enhanced pre-existing Th1 type immunity.

View Article: PubMed Central - PubMed

Affiliation: Tumor Immunology Unit, San Raffaele Scientific Institute, Milan, Italy.

ABSTRACT

Background: Pancreatic cancer is a very aggressive disease with dismal prognosis; peculiar is the tumor microenvironment characterized by an extensive fibrotic stroma, which favors rapid tumor progression. We previously reported that pancreatic cancer patients have a selective Th2 skew in the anti-carcinoembryonic antigen (CEA) CD4(+) T cell immunity, which correlates with the presence of a predominant GATA-3(+) tumor lymphoid infiltrate. This has negative effects in both effective anti-tumor immunity and further favoring fibrinogenesis. Aim of this study was to evaluate whether the Th2 polarization of CEA-specific CD4(+) T cells from pancreatic cancer patients is stable or can be reverted by immunomodulating cytokines.

Methodology/principal findings: We first evaluated the influence of IL-12 and IL-27, as single agents and in association, on the polarization of CEA-specific Th2 CD4(+) T cell clones from a pancreatic cancer patient. We found that only the combination of IL-12 and IL-27 modified the polarization of Th2 effectors by both reduction of IL-5, GM-CSF and IL-13 and induction of IFN-gamma production, which lasted after cytokine removal. Second, we evaluated the effect of the combined treatment on polyclonal CEA-specific CD4(+) T cells in short-time re-stimulation assays. In agreement with the data obtained with the clones, we found that the combined treatment functionally modulated the Th2 polarization of CEA-specific CD4(+) T cells and enhanced pre-existing Th1 type immunity.

Conclusions/significance: Collectively, our results demonstrate that tumor antigen specific Th2 CD4(+) T cells in pancreatic cancer are endowed with functional plasticity. Hence, loco-regional cytokines delivery or targeted therapy based on antibodies or molecules directed to the tumor stroma might improve anti-tumor immunity and ameliorate fibrosis, without systemic toxicity.

Show MeSH
Related in: MedlinePlus