Enhancing Anti-Tumor Immunity via Nrp1 Blockade

On day 1, C57BL/6 mice were immunized with 10⁶ irradiated (6,000 rad) B16-OVA cells i.p. in 200 μl PBS. Seven days later, mice were given high-dose gp96 i.d. in 100 μl PBS (or PBS only as control). On the same day, mice also received 100 μg Nrp1 blocking antibody (clone 761704, RnD Systems) or 100 μg rat IgG2a isotype antibody (clone eBR2a, eBioscience) via the i.p. route. Route and dose of Nrp1 antibody is based on previous reports using this antibody38 (link). One week later, all mice were killed and draining lymph nodes were harvested for flow cytometry. The percent of Foxp3+ T cells was quantified.