Anti il 9

Acute Model: Wild type (WT) and Sfpi1^lck−/− mice were sensitized by intraperitoneal injection of OVA (Sigma) adsorbed with alum (Sigma) on days 0 and 7 and subsequently challenged with intranasal OVA for 5 days as described previously (5 (link)). Where specified, mice were given 20 μg control antibody or anti-IL-9 (222622; R&D Systems) intravenously 30 min before the first, third and fifth challenges. Mice were sacrificed 48 h after the final intranasal challenge. Chronic model: WT and Sfpi1^lck−/− mice were sensitized by intranasal injection of 40 μg HDM extract (Dermatophagoides pteronyssinus in phosphate-buffered saline, PBS) from Greer Laboratories (Lenoir, NC) or PBS 3 days per week for 5 weeks. Mice were sacrificed 24 h after the final intranasal challenge. Cells from mediastinal lymph nodes were stimulated with HDM for 5 days, and cytokine production measured by ELISA.

Briefly, differentiated OVA-specific Th2 or Th9 cells were adoptively transferred intravenously into wild-type recipient mice (33 (link)). Twenty-four hours after cell transfer, mice were challenged intranasally with 100 μg OVA plus 500 ng TSLP for 5 days. Mice were then sacrificed 24 h after the last challenge for further analysis. To neutralize cytokine in recipients of Th2 or Th9 cells, we injected mice via tail vein with anti-IL-9 (10 μg/dose), anti-IL-13 (10 μg/dose), or IgG2b control Ab (10 μg/dose, R&D Systems) on days 1, 3, and 5.

Inhibitors were purchased from commercial suppliers as listed below, or were provided (Compound 29, (Patch et al., 2011 (link))) by the Donald McDonnell Laboratory (Duke University). ABT199 (Selleckchem #S8048), ABT263 (Selleckchem #S1001), LCL161 (Selleckchem #S7009), Birinapant (Selleckchem #S7015), Fomepizole (Selleckchem #S1717), CHR2797 (Tocris #3595), XCT790 (Sigma Aldrich #X4753), Topiramate (Selleckchem #S1438), ACET (Tocris #2728), Almorexant (Selleckchem #S2160), SB334867 (R&D Systems, #1960), anti-IL1R1 (R&D Systems, #AF269), anti-IL9 (R&D Systems, #AF209), anti-ITGAV (Abcam, #ab16821), Cilengitide (Selleckchem, #S7077), Guanidine HCl (Sigma Aldrich, #G3272), Dalfampridine (Sigma Aldrich, #275875), BEZ235 (Selleckchem, #S1009), Apitolisib (Selleckchem, #S2696), S17092 (Sigma Aldrich, #SML0181), WNK463 (Selleckchem, #S8358).