Recombinant il 22

In some experiments, pancreatic islet allograft recipient mice also received anti-NK1.1 or isotype control antibodies (BioXcell, clone PK136, 0.5 mg per mouse) 24 h before and 0 and 24 h after transplantation via a tail vein injection60 (link). In other experiments, some of the pancreatic islet allograft recipient mice also received anti-IL-22 (eBioscience, clone JES6-1A12, 0.3 mg per mouse), anti-NKG2A (Bio-Rad, clone 20D5, 0.3 mg per mouse)22 (link)23 or isotype control antibodies (Rat IgG2a K isotype control antibody, eBioscience, Clone: eBR2a, 0.3 mg per mouse) 24 h before and 0 and 24 h after transplantation via a tail vein injection.
Recombinant IL-22 (BioLegend) (100 ng g⁻¹ body weight) or normal saline was administered to some allograft recipient mice via an intravenous injection. For in vitro experiments, islets were cultured in the presence of Recombinant IL-22 or IFN-γ (BioLegend) at a concentration of 10–20 ng ml⁻¹ for 24 and 48 h.

In some experiments, mice were treated with recombinant IL-22. One month after the induction of T2DM, mice were challenged with aerosolized Mtb. Five months p.i., one group of mice were intravenously treated with a single dose of recombinant IL-22 (100 ng/kg body weight) and another group of mice was treated with recombinant IL-22 (100 ng/kg body weight) (BioLegend) or PBS twice a week.

The mouse small intestinal epithelial cell line, IEC4, was maintained by serial passage in DMEM medium supplemented with HEPES (1%), L-glutamine (10%), Pen/Strep (1%) and FBS (5%) (all from Gibco, USA). One-million IEC4 cells were seeded in 6-well plates and cultured for 48 hr. Scolices and 2 cm of strobila of H. diminuta retrieved from the small intestine of rats or IL-4 receptor-α^-/- mice (fail to expel H. diminuta) were exposed to a cocktail of antibiotics (Gentamicin solution, Sigma, St. Louis, Mo, USA) for 2 hr. A single worm was added to IEC4 monolayers ± recombinant IL-22 (5 ng/ml; Biolegend, CA, USA), and supernatants collected for measurement of IL-25 and then total RNA extracted.