Dorsal air pouches were prepared by injection of 2.5 mL of air on day 0 and day 3 in CD-1 mice, as previously described.25 (link) On day 6, mice received 0.25 mL of one of the following treatments diluted in 0.5% carboxymethyl cellulose (CMC, Sigma-Aldrich): (1) vehicle, CMC alone; (2) IL-17 (1 µg); (3) nIL-17 (1 µg); (4) IL-17 (1 µg) plus MAB421 or Ab-IPL-IL-17 (10 µg); (5) IL-17 (1 µg) plus anti-JE (10 µg, MAB479, R&D System) and (6) IL-17 (1 µg) plus anti-KC (10 µg, MAB453, R&D System). Mice were sacrificed after 24 hours, and lavage fluids were recovered, and centrifuged at 220 g for 10 min at 4°C. Cell pellets and inflammatory exudates were banked for subsequent analysis of inflammatory cyto-chemokines. The route, timing and frequency of administration as well as the selected dosages of tested compounds were selected according to updated literature.12 16 (link) Cell number was determined by TC20 automated cell counter (Bio-Rad) using Bio-Rad’s TC20 automated cell counter uses disposable slides, TC20 trypan blue dye (0.4% trypan blue dye w/v in 0.81% sodium chloride and 0.06% potassium phosphate dibasic solution, Sigma-Aldrich) and a CCD camera to count cells based on the analyses of capture images.25 (link)
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