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Inhibition of the Th2 response potentiates IL-17A and IL-17F production but is not sufficient to exacerbate neutrophil recruitment or AHR development.

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posted on 19.09.2013, 02:10 by Rebecca A. Martin, Jennifer L. Ather, Rebecca Daggett, Laura Hoyt, John F. Alcorn, Benjamin T. Suratt, Daniel J. Weiss, Lennart K. A. Lundblad, Matthew E. Poynter

Mice were exposed to 15ppm NO2 for 1 hour on day 1. All mice were exposed to OVA on days 1-3 and again during challenge on days 14-16 and analyzed 48 hours after the final antigen challenge. IgG isotype control antibody was administered one day prior to NO2 exposure and one day prior to OVA challenge. Anti IL-4 neutralizing antibody was administered one day prior to sensitization on day 0. A second group also received anti-IL-4 antibody one day prior to OVA challenge on day 13. Because we observed no differences in AHR, BAL cell counts, or cytokine production between mice that received anti-IL-4 at sensitization only on day 0 or on day 0 and day 13, we combined anti-IL-4 treated mice for statistical analysis. 48 hours following the final antigen challenge, lungs were removed and enzymatically digested. Single cell suspensions were restimulated in the presence of 200 μg/ml OVA antigen for 96 hours prior to the harvesting of cell supernatants and analysis of cytokine production. IL-17A (A) and IL-17F (B) were quantitated by Milliplex. IL-5 (B) and IL-13 (C) were quantified by ELISA. BAL total cells (E), Macs (F), PMNs (G), and Eos (H) were determined. Percent baseline and average percent baseline per dose of methacholine were calculated for R (I-J) and E (K-L). Statistics were performed by student’s t-test (A-D), 2-way ANOVA (J and L) or 1-way ANOVA (E-H) and Tukey post hoc analysis. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 compared to non-inflamed unless otherwise indicated by brackets. ND, not significantly different compared to non-inflamed. n = 6-12 per group.