To induce pancreatitis, mice aged 4–6 weeks were treated with 8-hourly intraperitoneal (i.p.) injections of caerulein (Sigma-Aldrich) at a dosage of 75 µg/kg body weight over 2 consecutive days. DT (Enzo Life Science) was administered i.p. every 4 days at a concentration of 25 ng/g. To establish the subcutaneous tumour model, 2×106 of primary mouse pancreatic cancer cell lines iKras*1, iKras*2 and iKras*3 derived from iKras*p53* tumours,11 (link)
13 (link) 4×105 of 65 671 cells (FVB/NJ strain)14 (link) or 7940B cells (C57BL/6J strain)15 (link) derived from KPC tumour (P48-Cre; loxP-stop-loxP (LSL)-KrasG12D; p53flox/+) were injected into CD11d-DTR mice of compatible genetic background. Tumour diameters were measured with digital callipers and the tumour volume was calculated by the formula: length×width2/2. For CD8+ T-cell depletion, anti-CD8 monoclonal antibody (BioXcell #BE0061; clone 2.43; 200 µg/mouse) was injected i.p. twice per week. Purified anti-mPD-1 antibody (BioXcell #BE0033-2; clone J43) was used for in vivo PD-1 blockade at a dose of 200 μg/i.p. injection, repeated every 3 days if needed. MEK inhibitor (MEKi) GSK1120212 (Selleckchem) was administered daily (1 mg/kg, i.p.) in a 10% (2-Hydroxypropyl)-β-cyclodextrin solution.
13 (link) 4×105 of 65 671 cells (FVB/NJ strain)14 (link) or 7940B cells (C57BL/6J strain)15 (link) derived from KPC tumour (P48-Cre; loxP-stop-loxP (LSL)-KrasG12D; p53flox/+) were injected into CD11d-DTR mice of compatible genetic background. Tumour diameters were measured with digital callipers and the tumour volume was calculated by the formula: length×width2/2. For CD8+ T-cell depletion, anti-CD8 monoclonal antibody (BioXcell #BE0061; clone 2.43; 200 µg/mouse) was injected i.p. twice per week. Purified anti-mPD-1 antibody (BioXcell #BE0033-2; clone J43) was used for in vivo PD-1 blockade at a dose of 200 μg/i.p. injection, repeated every 3 days if needed. MEK inhibitor (MEKi) GSK1120212 (Selleckchem) was administered daily (1 mg/kg, i.p.) in a 10% (2-Hydroxypropyl)-β-cyclodextrin solution.
