Anti cd8 clone yts 169

Cyclophosphamide (Endoxan) was obtained from the Sir Charles Gardiner Hospital pharmacy (Perth, Australia) and dosed at 200 mg/kg. Chemotherapy drugs were prepared under sterile conditions and diluted in 0.9% saline solution. Drugs were administered intraperitoneally (i.p) at a maximum volume of 100 µl per 10 g weight of mouse.
All-trans-retinoic acid (≥ 98% by HPLC, powder) was purchased from Sigma-Aldrich (Macquarie Park, NSW) and suspended in dimethyl sulfoxide (DMSO; Sigma-Aldrich) at a concentration of 40 mg/ml for short term storage at − 80˚C. Mice were treated with 10 mg/kg of tretinoin for 9 consecutive days, intraperitoneally. Working concentrations of tretinoin were suspended in PBS and had a final DMSO concentration of 5%.
For all immune cell depletion experiments, antibody administration began three days before tumour inoculation. Anti-CD4 (clone GK1.5) and anti-CD8 (clone YTS 169; both BioXcell, New Hampshire, USA) were dosed i.p at 100 µg in 100 µl of PBS on day 3, day 0 and thereafter weekly until the end of experiment. Depletion was monitored in peripheral blood collected from the tail vein and analysed by flow cytometry.

Chemotherapy and ICB were administered on the same day, initiating treatment when tumors were approximately 20-25 mm² in size. Chemotherapies were provided by Sir Charles Gardiner Pharmacy (Nedlands, WA, Australia) and was administered at the predetermined MTD as previously reported (16 (link)), except 5-FU which was administered at 75 mg/kg because MTD 5-FU with ICB caused severe toxicity (Table S1). Anti-CTLA-4 (clone 9H10, JJP Biologics) was dosed once at 100 μg/mouse and anti‐PD‐L1 (clone MIH5, JJP Biologics) was dosed 3 times with 2-day intervals at 100 μg/mouse (17 (link)). For depletion experiments, anti-CD4 (clone GK1.5, BioXcell), anti-CD8 (clone YTS 169, BioXcell) or anti-IL1β (clone B122, BioXcell) antibodies were administered 3 times with 3-day intervals at 100 μg/mouse with the first dose commencing 3 days before chemo‐immunotherapy. Anti-TNFα (clone XT3.11, BioXcell) was administered using the above schedule but at 2 mg/mouse. All treatments were diluted in sterile 0.9 % sodium chloride and administered intraperitoneally (i.p.) or intravenously (i.v.) as described in Table S1.