Figure 6. Tmem173−/−Mice Show Defective Accumulation of Antitumor T Cells.
(A and B) CFSE-labeled 2CT cell were transferred into WT or Tmem 173−/− mice and B16.SIY melanoma cells were inoculated into recipient mice after 1 day. On day 6, mice were sacrificed and spleens and tumor-draining lymph nodes were removed. Cells were stained with anti-CD8 and the clonotypic mAb 1B2 and analyzed by flow cytometry for CFSE dilution. Data were analyzed by FlowJo software and quantitated as shown in the panels on the right.
(C) BM-DCs from WT or Tmem173−/− mice were stimulated with tumor DNA for 7 hr and RNA was isolated. Isolated RNA was analyzed by Affymetrix GeneChip analysis.
(D-F) BM-DCs from WT or Tmem173−/− mice were stimulated with tumor DNA and the indicated cytokines were measured by ELISA.
(G) Mice were injected with 1 × 106 B16-SIY cells on day 0. On day 4, combinatorial antibody therapy was initiated. Anti-CTLA-4 mAb (100 ng/mouse) was administered intraperitoneally on days 4,7, and 10, and anti-PD-L1 mAb (100 ng/mouse) was given every other day starting on day 4 and ending on day 16. Tumor growth was measured on the indicated days. Data represent mean ± SEM (n = 10, each group) of combined two independent experiments. Statistical significance was determined using two-way ANOVA test. *p < 0.05, **p< 0.01, ***p < 0.001, ****p < 0.0001 (Student’s t test or two-way ANOVA test). Data indicate mean ± SEM (n = 5) and representative of two (A and B) or three (C-F) independent experiments.