LSECtin, expressed by B16 cells, inhibits the tumor-specific immune responses both in vivo and in vitro. LSECtin, expressed by B16 cells, inhibits the tumor-specific immune responses both in vivo and in vitro. A, WT C57BL/6 mice were inoculated with B16-LSECtin and B16-mock cells, and the TDLN cells or splenocytes were isolated for IFNγ intracellular staining and ELISPOT assays. B, representative dot plots for intracellular IFNγ staining and the percentages of IFNγ+ T cells in total CD8+ T cells are shown. *, P < 0.05. C and D, IFNγ ELISPOT assays of TDLN cells (C) and splenocytes (D) from mice inoculated with B16-LSECtin (black) and B16-mock (gray) cells in the presence or absence of TRP2 180–188. E, WT C57BL/6 mice were inoculated with irradiated B16 cells, and TDLN cells or splenocytes were isolated for coculture assays. F, TDLN cells (left) or CD8+ T cells (right) were subjected to restimulation ex vivo with irradiated B16-LSECtin (black) and B16-mock (gray) cells. *, P < 0.05; **, P < 0.01. G, CD4+ T cells (CD4+CD25−) or Treg-enriched cells (CD4+CD25+), sorted from restimulated splenocytes, were also subjected to restimulation ex vivo with irradiated B16-LSECtin (black) and B16-mock (gray) cells in the presence of 2 μg/mL coated anti-CD3. **, P < 0.01. These results are representative of three independent experiments. Feng Xu et al. Cancer Res 2014;74:3418-3428 ©2014 by American Association for Cancer Research