Anti pd 1 mab clone rmp1 14

All animal experiments were approved by the Institutional Animal Care and Use Committee of CHA University and KLSbio. The mice used here were maintained and handled according to policies approved by CHA University and KLSbio, respectively. Five-week-old female C57BL/6 mice were purchased from Orient Bio (Gyeonggi, Korea). For the syngeneic tumor mouse model, a total of 2 × 10⁵ MC38 cells were subcutaneously inoculated in C57BL/6 mice. The mice were orally administered the L. lactis strain daily and were intra-peritoneally treated with anti-PD-1 mAb (clone RMP1-14; BioXCell, Lebanon, NH, USA) or 3 mg/kg oxaliplatin (S1224, Selleckchem, Houston, TX, USA) on days 1, 4, 8, 11, and 14. The tumor size was monitored three times per week until the study endpoint. The tumor volume was calculated as the length × width² × 0.5.
Nude mice were purchased from Orient Bio (Gyeonggi, Korea). Nude mice were orally administered the L. lactis strain for 14 days. Then, a total of 2 × 10⁵ CT26, 1 × 10⁶ 4T1, and 1 × 10⁶ HCT116 cells were inoculated subcutaneously into the flank of each mouse. The nude tumor mice were administered the L. lactis daily, and the tumor size was monitored three times a week until the study endpoint. The tumor volume was calculated as the length × width² × 0.5.

All animal experiments were performed with approval from the Institutional Animal Care and Use Committee of CHA University (permission number IACIC180010). Mice used in this study were maintained and handled according to policies approved by CHA University. Five-week-old female C57B6/N mice were purchased from the Orient Bio (Gapyeong, Gyeonggi, Korea). For the tumor mice model, mice were orally administered Bifidobacterium strains for 14 days (−14 day). Then, mice were implanted with 2 × 10⁵ MC38 colon adenocarcinoma cells subcutaneously and treated with 3 mg/kg oxaliplatin (S1224, Selleckchem, Houston, TX, USA) or 2 mg/kg anti-PD-1 mAb (clone RMP1-14, BioXCell, USA) intraperitoneally. Syngeneic tumor mice were treated 6 times with anti-PD-1 mAb or oxaliplatin on days 3, 7, 10, 14, 17, and 21. Tumor size was monitored three times a week until the study endpoint. Tumor volume was calculated as length × width² × 0.5.

Mice were subcutaneously injected into the right flank with 5 × 10⁵ tumor cell line (n = 5 per group). When tumors reached a size of 20 to 40 mm³, mice were then injected intraperitoneally (i.p) at 3-day intervals with 250 μg of anti-PD-1 mAb (clone RMP1–14, BioXcell, NH, USA) or isotype control (clone MPC11). The experimental group were the following: 1) Isotype, 2) anti-PD-1 treatment, 3) Isotype and CBM588, 4) anti-PD-1 treatment and CBM588. Tumor growth over two-weeks were routinely monitored by means of a caliper. Briefly, measurements in mm of two perpendicular diameters using a Caliper were used every 2–3 days as a metric of subcutaneous tumor volume for monitoring tumor growth calculated according to the formula largest diameter multiplied with smallest diameter²/2. Mice were classified as responder when the specific growth rate (% increased volume at day 14 when compared at day 7 before initiation of PD-1 blockade) of their tumor was decreased by more than 40% when compared to the median of specific growth rate of tumors from the group of mice that received the isotype. Experiments were performed at least twice.