Journal: The Journal of Clinical Investigation

Article Title: T cell–intrinsic VISTA expression promotes resistance to CTLA-4 blockade by restricting CD8 + T cell responses

doi: 10.1172/JCI195668

Figure Lengend Snippet: ( A ) Mice were inoculated on the flank with murine tumors, including B16bl6 melanoma (100,000), E0771 breast cancer (100,000), MC38 colon cancer (100,000), and LLC lung cancer cells (100,000). Tumors were harvested when reaching diameters of approximately 7–8 mm, and TILs were examined for VISTA expression. At least 2 tumors were analyzed for each tumor type. Representative results are shown. ( B ) VISTA expression was quantified by MFI on CD8 + TILs from B16.OVA tumors, including the PD-1 + and PD-1 + CD103 + subsets. * P < 0.05, *** P < 0.001, and **** P < 0.0001. Significance was determined by 1-way ANOVA. Data are presented as mean ± SEM. ( C ) VISTA expression on CD8 + TILs from surgically resected human tumor tissues was analyzed by flow cytometry. The data presented include CD8 + TILs from melanoma tissues, head and neck tumors, endometrial tumors, and breast cancer tissues. When available, multiple patient (Pt) specimens were included in the analysis.

Article Snippet: B16bl6, B16.OVA, MC38 (Sigma-Aldrich, catalog SCC172), LLC (ATCC, CRL-1642), and E0771.lmb (ATCC, CRL-3405) murine tumor cell lines were cultured in bicarbonate-free, HEPES-free RPMI-1640 supplemented with 10 mM HEPES (Life Technologies), 10% FBS, sodium pyruvate, 2 mM l -glutamine, 50 μM β-mercaptoethanol, and penicillin/streptomycin (Life Technologies).

Techniques: Expressing, Flow Cytometry