Tryptase and cytokines levels in peritoneal lavage
For assessing tryptase and
cytokine levels in the peritoneal cavity, OVA/SEB AP-induced and
challenged mice were euthanized 30 min or 48h after challenge
respectively and peritoneal lavages (1 ml) were collected. Cells were
separated from supernatants by centrifugation and supernatants were
cleared by any remaining cell by a second centrifugation step. Tryptase
levels were assayed as described before (15). Cytokines levels were
determined by using IL-4 (900-K49; PeproTech, Rehovot, IL), INFÉ£
(900-K98; PeproTech) and IL-17A (900-K392; PeproTech) ELISA kits
according to the manufacturer’s instructions.