Anti cd279

CD8⁺T cells isolated from human PBMC were cultured in the presence of plate-bound anti-CD3 (1 µg/ml) with different dosage VEGF-A (0, 20, 50ng/ml; R&D Systems). Cells were harvested and analyzed by flow cytometry after 84 h or used to extract mRNA after 60 h. T cells isolated from tumor of tumor-bearing mice were treated by the combination of antibody Anti-CD279 (BioXCell, BE0146) or IgG2a (BioXCell, BE0089) with different dosage anti-VEGFR2 antibody (DC101, BioXCell, BE0060) or IgG1 (BioXCell, BE0088). After 48 h, cells were analyzed by flow cytometry.

6–8 weeks old, male C57BL/6 mice were purchased from Shanghai Jihui Experimental Animal Breeding Co., Ltd. (Shanghai, China). Animals were housed and maintained under optimal conditions of light, temperature, and humidity with free access to food and water. For subcutaneous tumor model, a total of 1 × 10⁵ LA795 cells were resuspended in 200 µL PBS and inoculated subcutaneously into the right flank of mice. Tumor dimensions were measured by caliper every other day, and tumor volume (mm³) was estimated using the formula: tumor volume = (length) × (width)² × π/6. Different doses of anti-VEGFR2 antibody (DC101, BioXCell, BE0060) and IgG1 (BioXCell, BE0088) treatment were initiated 11 days after tumor cells inoculation and administered by intraperitoneal injection every 3 days. Anti-PD1 antibody (Anti-CD279, BioXCell, BE0146) and IgG2a (BioXCell, BE0089) treatment were initiated 12 days after tumor cells inoculation and was administered at 200 µg/mouse by intraperitoneal injection every 3 days. At indicated days later, the tumor-bearing mice were anesthetized and tissues were harvested for further analysis and measurement. Survival analysis was continued as independent experiments for indicated days.