Th17-polarized TRP-1 cells have a survival advantage after adoptive transfer into tumor-bearing hosts. (A) Spleens, lymph nodes, and tumors were harvested on day 6 from nontreated animals (NT) or animals treated with Th0, Th1, or Th17 cells and analyzed by flow cytometry for expression of Vβ14 and CD4. The panel is representative of 4 distinct experiments. (B) The total number of Vβ14⁺CD4⁺ cells recovered from spleens of treated animals on days 6 and 12 was calculated as described in “Methods” (± SD, n = 3-4). (C) In vivo proliferation of polarized TRP-1 cells. CFSE-labeled Th0, Th1, or Th17 (Thy1.2⁺) cells were adoptively transferred into 500 R irradiated B6.PL hosts (Thy1.1⁺). Splenocytes were harvested on days 3 and day 6 and analyzed by flow cytometry. Histograms show CFSE fluorescence after gating on Thy1.2⁺ population. Th0 and Th1 displayed a greater dilution of the florescent dye compared with Th17 cells. (D) Polarized TRP-1 T cells (Thy1.2⁺) were transferred into sublethally irradiated tumor-bearing B6.PL (Thy1.1⁺) hosts. The frequency of Th1.2⁺CD4⁺ cells in tumor-draining lymph nodes pooled from at least 3 animals/group was measured by flow cytometry 6 days after adoptive cell transfer.