Ty, and no sex-specific variations were observed. Groups in individual experiments have been sex-matched and age-matched. All animals were housed beneath specific-pathogen-free circumstances in the National Institutes of Health in an American Association for the Accreditation of Laboratory Animal Careapproved facility. S. mansoni egg nduced lung granuloma model five,000 S. mansoni eggs (Biomedical Investigation Institute) have been P2X1 Receptor Antagonist web injected intraperitoneally on day 0 to sensitize mice. On day 14, mice were challenged once again intravenously with 5,000 reside eggs containing mature embryos before lungs were Traditional Cytotoxic Agents Inhibitor Biological Activity harvested on day 18 or 21. Schistosome egg antigen nduced lung inflammation model Schistosome egg antigen (SEA) was obtained from sterile LPS-free liver-derived eggs from mice chronically infected with Schistosoma mansoni. Mice were intratracheally sensitized and challenged with 10 SEA on days 0, 7, 14, 16, and 18. SEA was administered in 30 saline to mice anesthetized with isoflurane. Lungs have been lavaged and harvested for analysis on day 19.Author Manuscript Author Manuscript Author ManuscriptAcute house dust mite allergen nduced lung inflammation model Mice were intranasally sensitized with 25 of home dust mite (HDM, Greer) or PBS on days 0, 1, and two. On days 15, 16, 17, and 18, mice have been intranasally challenged with 5 of HDM or PBS. HDM was administered in 30 PBS to mice anesthetized with isoflurane. Lungs had been lavaged and harvested for evaluation on day 19.Nat Immunol. Author manuscript; available in PMC 2017 Could 01.Vannella et al.PageChronic residence dust mite allergen nduced lung inflammation modelAuthor Manuscript Author Manuscript Author Manuscript Author ManuscriptMice had been intranasally sensitized with 25 of house dust mite (HDM, Greer) or PBS on days 0, 1, and two. On days 14, 15, 16, 28, 29, 30, 42, 43, and 44, mice have been intranasally challenged with five of HDM or PBS. Lungs were lavaged and harvested for evaluation on day 45. Acute papain-induced lung inflammation model Mice have been intranasally sensitized with 12.5 of papain (Carica papaya, EMD Millipore) or water on days 0, 1, and 2. On days 15, 16, 17, and 18, mice have been intranasally challenged with ten of papain or PBS. Papain was administered in 30 PBS to mice anesthetized with isoflurane. Lungs were lavaged and harvested for evaluation on day 19. Nippostrongylus brasiliensis infection Third-stage (L3) larvae have been ready as described previously31, and 500 were injected subcutaneously in to the nape in the neck of every mouse. Feces were collected on days 60 post-infection for egg counts. Adult worms have been counted in the small intestine on days five, 8, 10, or 14. Representative sections of lung and small intestinal tissue were taken for histology and qPCR evaluation on day eight. Heligmosomoides polygyrus bakeri infection As described previously32, mice were inoculated periorally with 200 L3, and 2 weeks later, worms have been expelled by administering 1 mg pyrantel pamoate. 4 weeks later, the sensitized mice had been challenged with H. p. bakeri (secondary) whilst naive mice have been inoculated with 200 L3 for the first time as controls (main). 12 d after this, mice have been sacrificed for evaluation. Eggs were counted in the feces, and tissue was collected for histology and gene expression assay by qPCR. Adult worms had been counted in the intestines 15 d right after principal inoculation and secondary challenge. The ATPLite Luminescence Assay Program (PerkinElmer) was applied to measure ATP content of adult worms collected from the i.
