c-Rel expression in donor T cells is increased after allo-HSCT.

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c-Rel expression in donor T cells is increased after allo-HSCT. c-Rel expression in donor T cells is increased after allo-HSCT. A, sublethally or nonirradiated BALB/c recipients were transplanted with C57BL/6 T cell depleted (TCD) BM cells with or without 1 × 106 C57BL/6 wild-type (WT) T cells. Levels of IL-2, CD25, and c-Rel are shown after gating on either donor or residual recipient T-cell populations on day 7. Values, mean ± SEM (n = 5). XRT, irradiation; *, P < 0.05; **, P < 0.01; MFI, mean fluorescence intensity. B and C, splenocytes were stimulated with phorbol 12-myristate 13-acetate (PMA)/ionomycin for 5 hours and analyzed for intracellular and intranuclear c-Rel expression by flow cytometry. B, intracellular c-Rel staining was performed following fixation and permeabilization (eBioscience kit). C, cytoplasm was removed from c-Rel–stained cells by 0.03% saponin with nuclear isolation media (NP-40, nonyl phenoxypolyethoxylethanol) to confirm nuclear localization of c-Rel. D, lethally irradiated BALB/c recipients were transplanted with C57BL/6 WT or c-Rel−/− TCD BM cells. Thymus, BM, and spleen were analyzed on day 28 after HSCT. Values, mean ± SEM (n = 4). Data shown in A to D are representative of two independent experiments. DC, dendritic cells; MDSCs, myeloid-derived suppressor cells; NK, natural killer. E, lethally irradiated BALB/c recipients were transplanted with C57BL/6 WT or c-Rel−/− TCD BM cells with or without 1 × 106 C57BL/6 WT or c-Rel−/− T cells. Survival curve is shown. F, lethally irradiated C57BL/6 WT or c-Rel−/− recipients were transplanted with B10.BR TCD BM cells with or without 2 × 106 B10.BR T cells. Survival curve is shown. Data in E and F are combined from three independent experiments, and values represent mean ± SEM (n = 21). ***, P < 0.001; n.s., not significant. Yusuke Shono et al. Cancer Discovery 2014;4:578-591 ©2014 by American Association for Cancer Research