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Figure 6 | Journal of Translational Medicine

Figure 6

From: Decitabine immunosensitizes human gliomas to NY-ESO-1 specific T lymphocyte targeting through the Fas/Fas Ligand pathway

Figure 6

Decitabine sensitizes glioma cells to an immune-mediated cell death via the Fas/FasL pathway. A) Decitabine-treated and untreated T98G cells were stained with an anti-Fas antibody and analyzed by flow cytometric analysis. B) T98G cells were treated as described previously and stained with an agonistic-Fas antibody (clone CH-11) to stimulate apoptosis for 24 hr. Results depict the average percentage of late cell death in untreated alone, decitabine-treated alone, untreated with agonistic mAb, and decitabine-treated with agonistic mAb. Decitabine-treated alone versus decitabine-treated with antibody (P = 0.0049). Untreated alone versus untreated with antibody (P = 0.0215). Experiments were conducted in triplicate. C) Representative flow cytometric analysis of CD107A staining from NY-ESO-1 TCR transduced T cells co-cultured with untreated T98G glioma cells, with or without the addition of an antagonistic-Fas antibody (clone ZB4) to block apoptosis (solid filled, NY-ESO-1 specific T cells alone; solid line, NY-ESO-1 specific T cells co-cultured with untreated T98G glioma cells; dashed line, NY-ESO-1 specific T cells co-cultured with untreated T98G glioma cells with antagonistic-Fas antibody. NY-ESO-1 TCR transduced T cells were gated from the CD8 population. D) Representative flow cytometric analysis of CD107A staining from NY-ESO-1 transduced T cells co-cultured with decitabine-treated T98 glioma cells, with or without the addition of an antagonistic-Fas antibody (clone ZB4) to block apoptosis (solid filled, NY-ESO-1 specific T cells alone; solid line, NY-ESO-1 specific T cells co-cultured with decitabine-treated T98G glioma cells; dashed line, NY-ESO-1 specific T cells co-cultured with decitabine-treated T98G glioma cells with antagonistic-Fas antibody. E) CBA was performed on cell-free supernatant of samples from Figure 6C. Th1/Th2 cytokine concentrations IFN-γ, TNF-α, and IL-5 (pg/mL) are depicted. (*p < 0.05; ** p < 0.01; NS, not significant).

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