Abstract
Th17 cells, a recently discovered inflammatory T cell subtype, have been implicated with autoimmune disorders. However, mechanism of generation or functions of intratumoral Th17 cells are still unclear. We have been investigating the mechanism of induction and role of Th17 cells in malignant gliomas using primary tumor as well as cell lines. We report here that: (1) a higher frequency of Th17 cells in gliomas were associated with higher number of myeloid (CD11b) cells as well as the expression of TGF-β1 or IL-6; (2) conditioned medium from glioma cells (Gl CM) induced Th17 cell differentiation, which was inhibited by anti-TGF-β1 and anti-IL-6; (3) glioma-associated monocytes secreted Th17-promoting cytokines IL-1β and IL-23; (4) CM from glioma and monocyte co-culture (Gl+Mo CM) induced high frequency of Th17 cells in naïve T cell culture, which was abrogated by anti-IL-1β and anti-IL-23 antibodies; (5) In vitro Gl+Mo CM-mediated Th17 generation was associated with a decrease in IFN-γ and a concomitant increase in IL-10 secretion. Anti-TGF-β1, but not anti-IL-6, significantly reversed this cytokine profile. These results demonstrate prevalence of Th17 cells in gliomas and implicate the cytokines derived from the tumor as well as infiltrating myeloid cells in the induction of Th17 cells in glioma microenvironment. Moreover, the data also suggest that glioma-associated Th17 cells may contribute to immune-suppression via TGF-β1-induced IL-10 secretion. Further studies on the mechanism of tumor-infiltration, developmental pathways, and pro-/anti-tumor functions of Th17 cells will provide rationale for developing novel adjuvant immunotherapeutic strategies for malignant gliomas.
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Abbreviations
- CM:
-
Conditioned medium
- nT:
-
Naïve T cells
- GI-CM:
-
Conditioned medium from glioma cells
- Mo-CM:
-
Conditioned medium from monocyte culture
- TGF:
-
Transforming growth factor
- CCL:
-
Chemokine ligand
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Acknowledgments
We acknowledge research support from the Fund for Medical Research and Education (FMRE) to PP. The Microscopy, Imaging and Cytometry Resources Core is supported, in part, by NIH Center grant P30CA22453 to The Karmanos Cancer Institute, Wayne State University and the Perinatology Research Branch of the National Institutes of Child Health and Development, Wayne State University. We are thankful to Dr. Indrajit Sinha for his critical evaluation of the manuscript and also for his help with the preparation of the Figures.
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262_2012_1312_MOESM1_ESM.ppt
Supplementary Figure S1. Prevalence of Th17 cells in gliomas. Paraffinized tumor sections, obtained from patients with malignant glioma were analyzed by IHC to determine the presence of CD4+IL-17+ Th17 cells (A); Isotype antibody control staining were also performed to rule out the background (B). The micrographs were imaged at 200x magnification (PPT 1389 kb)
262_2012_1312_MOESM2_ESM.ppt
Supplementary Figure S2. Isotype Controls for data presented in Figure 6. nT cells were cultured, stained with isotype control antibodies and were analyzed by flow cytometry, as described under Figure 6. The data are from one representative experiment out of three experiments performed with similar results (PPT 294 kb)
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Paladugu, M., Thakur, A., Lum, L.G. et al. Generation and immunologic functions of Th17 cells in malignant gliomas. Cancer Immunol Immunother 62, 75–86 (2013). https://doi.org/10.1007/s00262-012-1312-7
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DOI: https://doi.org/10.1007/s00262-012-1312-7