Anti pd 1 antibody clone j43

Mice were given 250 µg anti-PD-1 antibody (clone J43, BioXCell) intraperitoneally every other day for 5 injections prior to harvest and compared to untreated controls.

Mice were monitored daily and tumor volume was measured with a calliper (volume = length x width² x 0.5). When tumors reached a suitable volume (approx. 70-100 mm³), mice were injected three times into the lateral tail vein with the pharmacological agents. Fusion proteins were dissolved in phosphate buffered saline (PBS), also used as negative control, and administered every 48h or 72h. The commercial anti-PD-1 antibody (clone J43, BioXCell) was administered i.v. once at a dose of 200 µg. For the tumor re-challenge study, mice with complete responses were injected subcutaneously with 5 x 10⁶ WEHI-164 cells in the flank.
For the in vivo depletion of CD4⁺ T cells, CD8⁺ T cells and NK cells, WEHI-164 tumor bearing mice were injected intra-peritoneally with 30 µL anti-Asialo GM1 (Wako 986-10001), 250 µg anti-CD4 (clone GK1.5 BioXCell) or 250 µg anti-CD8 (clone 2.43 BioXCell) antibodies on day 2, 5, 8 and 11 after tumor implantation. A saline group and a treatment group without depletion were included as controls.
Results are expressed as tumor volume in mm³ ± SEM. For WEHI-164 studies, 5 mice per group (6 mice per group for the depletion experiment) were used. For therapy studies with C1498, CT26 and F9 tumors, 4 mice per group were used.