Low-Dose Decitabine Augments the Activation and Anti-Tumor Immune Response of IFN-γ+ CD4+ T Cells Through Enhancing IκBα Degradation and NF-κB Activation

Abstract

Background CD4 + T cells play multiple roles in controlling tumor growth and increasing IFN-γ + T-helper 1 cell population could promote cell-mediated anti-tumor immune response. We have previously showed that low-dose DNA demethylating agent decitabine therapy promotes CD3 + T-cell proliferation and cytotoxicity; however, direct regulation of purified CD4 + T cells and the underlying mechanisms remain unclear. Methods The effects of low-dose decitabine on sorted CD4 + T cells were detected both in vitro and in vivo . The activation, proliferation, intracellular cytokine production and cytolysis activity of CD4 + T cells were analyzed by FACS and DELFIA time-resolved fluorescence assays. In vivo ubiquitination assay was performed to assess protein degradation. Moreover, phosphor-p65 and IκBα levels were detected in sorted CD4 + T cells from solid tumor patients with decitabine-based therapy. Results Low-dose decitabine treatment promoted the proliferation and activation of sorted CD4 + T cells, with increased frequency of IFN-γ + Th1 subset and enhanced cytolytic activity in vitro and in vivo . NF-κB inhibitor, BAY 11-7082, suppressed decitabine-induced CD4 + T cell proliferation and IFN-γ production. In terms of mechanism, low-dose decitabine augmented the expression of E3 ligase β-TrCP, promoted the ubiquitination and degradation of IκBα and resulted in NF-κB activation. Notably, we observed that in vitro low-dose decitabine treatment induced NF-κB activation in CD4 + T cells from patients with a response to decitabine-primed chemotherapy rather than those without a response. Conclusion These data suggest that low-dose decitabine potentiates CD4 + T cell anti-tumor immunity through enhancing IκBα degradation and therefore NF-κB activation and IFN-γ production.

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