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. 2017 Sep 22;7:12168. doi: 10.1038/s41598-017-12488-z

Figure 1.

Figure 1

Host pre-conditioning with CTX allows adoptively transferred tumor-specific CD4+ T cells to differentiate into polyfunctional effector cells and regain IL-7Rα expression. (A) Kinetics of IL-7Rα expression on donor CD4+ T cells following adoptive transfer. Following the timeline depicted in the schema, mice with established systemic A20HA tumors were divided into two groups. One group of mice were pre-conditioned with CTX while the other group received PBS. All mice received adoptive transfer of HA-specific CD4+ T cells the next day. At the indicated time points, tail blood samples were collected and analyzed for IL-7Rα expression on donor CD4+ T cells by FACS. IL-7Rα expression profiles relative to cell division in transferred CD4+ T cells are shown in representative dot plots. The numbers indicate the percentage of cells in the corresponding quadrant. Results are summarized in graph at right. Data for day 0 and day3 are gated on total donor T cells because cells barely divided at these two time points. (B) Differential expression of IL-7Rα in donor CD4+ T cells in mice with or without CTX pre-conditioning. 7 days after T-cell transfer, spleen cells were isolated to examine IL-7Rα expression on donor CD4+ T cells. Donor T cells were also evaluated for expression levels of PD1, Foxp3, CD40L, IFNγ and TNFα. The phenotypes of the divided donor T cells in PBS or CTX-conditioned mice are summarized in (C). (D) Donor CD4+ T cells transferred into CTX-conditioned mice but not unconditioned mice are capable of producing IL-2. IL-2 expression profiles relative to cell division in transferred CD4+ T cells are revealed by intracellular staining (ICS). (E) IL-2 neutralization inhibits IL-7Rα re-expression in donor CD4+ T cells in CTX-conditioned mice. Tumor-bearing mice were treated with CTX followed by CD4+ T cell transfer the next day. Some mice were injected with IL-2 neutralizing mAbs every other day. 7 days after T cell transfer, IL-7Rα expression on donor CD4+ T cells were analyzed by FACS. Results summarized in bar graph are shown as mean ± SD of 3 mice each group. (F) Exogenous IL-2 partially restores IL-7Rα expression in donor CD4+ T cells transferred into unconditioned mice. Following the timeline depicted in the schema, A20HA tumor-bearing mice received adoptive transfer of HA-specific CD4+ T cells. A cohort of mice further received IL-2/αIL-2 complex (IL-2C) every other day. IL-7Rα expressions on donor CD4+ T cells were analyzed 7 days after T cell transfer.