For Treg depletion, Foxp3DTR, KC;Foxp3DTR mice were treated with diphtheria toxin (DT) (50 ng/g) (Enzo Life Science) by intraperitoneal injection (i.p.). DT injections were repeated according to the specific experimental design shown in Figures. Control mice lacking Foxp3DTR alleles received the same DT treatments. In KC, KC;Foxp3DTR and KC;CD4−/− mice, mild acute pancreatitis was induced in 4–5-week-old mice by a series of 8 hourly intraperitoneal injections of caerulein (Sigma-Aldrich, 75 μg/kg) over 1-day period. For CCR1 inhibition, mice were subcutaneously dosed with CCR1 antagonist BX471 (Sigma-Aldrich, 50 mg/kg) for 7 days at 12-hour intervals. DMSO was used as vehicle to dissolve BX471 at a concentration of 50 mg/ml.
To establish the orthotopic pancreatic cancer model, 1×105 of 7940B cells (C57BL/6J strain)(46 (link)) derived from KPC tumor (Ptf1a-Cre; LSL-KrasG12D; p53flox/+) were injected into Foxp3DTR mice of compatible genetic background. Cells were tested for mycoplasma free by MycoAlertTM PLUS Mycoplasma Detection Kit (Lonza) and passage 15–20 were used for all experiments. For CD8+ T cell depletion, anti-CD8 mAb (BioXcell clone 2.43; 200 μg/mouse) was injected i.p. twice per week. For CD4+ T cell depletion, anti-CD4 mAb (BioXcell clone GK1.5; 200 μg/mouse) was injected i.p. at least every three days.
To establish the orthotopic pancreatic cancer model, 1×105 of 7940B cells (C57BL/6J strain)(46 (link)) derived from KPC tumor (Ptf1a-Cre; LSL-KrasG12D; p53flox/+) were injected into Foxp3DTR mice of compatible genetic background. Cells were tested for mycoplasma free by MycoAlertTM PLUS Mycoplasma Detection Kit (Lonza) and passage 15–20 were used for all experiments. For CD8+ T cell depletion, anti-CD8 mAb (BioXcell clone 2.43; 200 μg/mouse) was injected i.p. twice per week. For CD4+ T cell depletion, anti-CD4 mAb (BioXcell clone GK1.5; 200 μg/mouse) was injected i.p. at least every three days.
