Anti tnfα clone xt3

Mice were infected subcutaneously (s.c.) into the tail base with 2×10⁶ sfu R. typhi in 50 μl PBS. Either 1×10⁶ purified CD8⁺ or CD4⁺ T cells from naïve BALB/c, BALB/c IFNγ^-/- or BALB/c Perforin^-/- mice were adoptively transferred 1 day prior to infection. TNFα was neutralized by intraperitoneal application of 500 μg anti-TNFα (clone XT3.11, BioXCell, West Lebanon, US) in 200 μl PBS. Control mice received the same amount of isotype antibody (clone HRPN, BioXCell, West Lebanon, US). Treatment was performed every three days beginning on day 3 post infection. For the neutralization of IL-17A, anti-IL-17A (clone 13F3) was used (BioXCell, West Lebanon, US). 500 μg of the antibody were injected i.p. in 200 μl PBS every 2 days starting on day 2 post infection. Control mice received the same amount of isotype antibody (clone MOPC-21, BioXCell, West Lebanon, US).

For neutralizing TNF-α signaling in vivo, we performed intraperitoneal injections of 500 µg anti-TNF-α-clone XT3.11 (#BE0058, BioX Cell) at 3 h after MCAO. For blockade of IFN-γ, mice were treated intraventricularly (i.c.v.) with 1 µg Ultra-LEAF™ Purified anti-mouse IFN-γ antibody (505834, BioLegend) at 12 h after MCAO. Similar amounts of rat IgG (#BE0088, clone HRPN, BioXcell or 400431, BioLegend) were delivered via corresponding route as isotype control antibodies.

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.