Stereotactic radiotherapy increases Tregs in tumors.

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Volume 23, Issue 11, Pages (June 2018)

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Stereotactic radiotherapy increases Tregs in tumors. Stereotactic radiotherapy increases Tregs in tumors. A, Experimental design. 5 × 105 B16/F10, 1.5 × 106 MC38, or 3 × 105 RENCA cells were implanted subcutaneously (s.c.) to the flank of wild-type C57BL/6J mice (in B16/F10 or MC38 experiments) or BALB/c mice (in RENCA experiment), respectively on day 0. Mice received 10 Gy of stereotactic radiotherapy (RT) via SARRP on day 7 (B16/F10), day 16 (RENCA), day 10 (MC38), respectively. Tumors, draining lymph nodes (DLN), and spleens were harvested 7 days after the radiation. B, Tumor growth curves of B16/F10-bearing mice as in A. Nonirradiated tumors (control) in black line and irradiated tumors (RT) in red line, respectively. C, The absolute number of TIL-Tregs per gram tumor weight in the B16/F10 model. D–F, Representative flow plots (D) and quantitative bar graph of % Foxp3+ cells (E) and MFI of Foxp3 (F) in CD4+ cells from TILs. N = 7–15 per group, repeated at least 4×. G, Immunohistochemistry of Foxp3 in tumors (B16/F10, RENCA, MC38), 20× and 40× magnification as indicated. Foxp3+ cells are stained red in B16/F10 (due to the brown pigment in melanoma), and brown in RENCA and MC38 tumors. Error bars, SEM; ***, P < 0.001; **, P < 0.01; *, P < 0.05, determined by two-way ANOVA (B) or unpaired Student t test (C, E and F). Yuki Muroyama et al. Cancer Immunol Res 2017;5:992-1004 ©2017 by American Association for Cancer Research