Leaf anti mouse rat il 1β antibody

For immunizations prior to tumor inoculation, mice were injected subcutaneously (s.c.) into the right flank with PBS (unimmunized), WTL alone at a concentration equivalent to 2.5×10⁵ tumor cells per mice or with WTL plus 10 μg/mice of LPS, or WTL plus LPS plus 65 μg/mice of oxPAPC or PGPC, all emulsified in incomplete Freud’s adjuvant (IFA). In some experiments, LPS is replaced by MPLA. 15 days post immunization, mice were challenged s.c. on the left flank with 3×10⁵ of viable B16OVA cells, or 5×10⁵ of viable MC38-OVA cells, as indicated. Tumor-free mice were re-challenged s.c. into the upper back with a lethal dose of 6×10⁵ viable B16OVA or 1×10⁶ of viable MC38-OVA cells, as indicated. When indicated, mice were given 100 μg of LEAF anti-mouse/rat IL-1β antibody (BioLegend) by intravenous (i.v.) injection for five consecutive days; starting two days before receiving the immunization, then on day 1, day 2, and day 3 post-immunization to ensure chronic depletion of circulating IL-1β. The size of the tumors was assessed in a blinded, coded fashion every two days and recorded as tumor area (length × width) using a caliper. Mice were sacrificed when tumors reached 2 cm³ or upon ulceration.