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. Author manuscript; available in PMC: 2016 Oct 15.
Published in final edited form as: J Immunol. 2015 Sep 16;195(8):4028–4037. doi: 10.4049/jimmunol.1501291

Figure 7.

Figure 7

BAp:I-Ab-specific Treg conversion depends on TGFβ1 and leukemic cell antigen presentation. A, B. Leukemic cells were gated as CD45.1+, GFP+ and were stained for cytokines. Shown is the median fluorescence intensity of IL10 (A) and TGFβ1 (B). Three independent experiments are shown, comparisons were made with Wilcoxan Matched-Pairs test (2 paired groups). C, D. Shown is the overlay of IL10 (C) and TGFβ1 (D) expression on BAp:I-Ab Tetramer−, CD44− CD4+ T cells (Grey), BAp:I-Ab Tetramer+, CD44+ CD4+ T cells (Dashed), and Leukemic cells (Black). Shown are 10 mice from three independent experiments, whose results were concatenated. E, F. BAp:I-Ab-specific T cells were gated on BAp:I-Ab Tetramer (APC)+, CD44+ and were stained for cytokines. Shown is the median fluorescence intensity of IL10 (E) and TGFβ1 (F). Three independent experiments are shown, comparisons were made with Wilcoxan Matched-Pairs test (2 paired groups). G. Mice were inoculated with MHC-II replete leukemia or MHC-II−/− leukemia as labeled, or treated with Isotype or anti-TGFβ antibody. BAp:I-Ab-specific Tregs were gated as previously described and the percent Tregs was analyzed. Results are from three independent experiments, Kruskal-Wallace and Dunns’ test used to establish significance. H. Mice were treated as in G, and BAp:I-Ab-specific T cell count was analyzed as previously described. Results are from three independent experiments, Kruskal-Wallace and Dunns’ test used to establish significance. I. BAp:I-Ab-specific T cells from mice inoculated with MHC-II−/− leukemia or MHC-II replete leukemia and harvested 14 days later were analyzed for Ly6C expression. Shown are three independent experiments, significance was established with Mann-Whitney U test. J. BAp:I-Ab-specific T cells from mice inoculated with leukemia and treated with anti-TGFβ or isotype control were analyzed for Ly6C expression 14 days later. Shown are three independent experiments, significance was established with Mann-Whitney U test. K. Mice inoculated with MHC-II−/− leukemia or MHC-II replete leukemia were aged until leukemia-induced morbidity and survival to morbidity was analyzed. P-values from Log-rank test; three independent experiments shown.