IL-17A production by lung cells from NO2-promoted and allergically sensitized mice is qualitatively different from IL-17A production by lung cells following Th17 adoptive transfer and antigen challenge.
Mice were subjected to NO2-promoted allergic sensitization, OVA-challenged, and analyzed 48 hours after the final antigen challenge. For adoptive transfer, CD4+ T-cells from OTII splenocytes by were either Th2 or Th17 polarized in vitro and adoptively transferred to recipient mice, which were then OVA-challenged for 3 consecutive days and analyzed 24 hours following the final OVA challenge. At analysis, lungs were removed and enzymatically digested. Lung single cell suspensions were restimulated with OVA antigen in the presence or absence of anakinra (Ana; 0.2 μg/mL), Dex (10-8 M), or anakinra and Dex in combination for 96 hours. Cell supernatants from antigen-restimulated and treated lung cells from mice subjected to NO2-promoted allergic sensitization and OVA challenge (A-D), Th17 adoptive transfer (E-H), or Th2 adoptive transfer (I-L) were harvested and cytokine levels were quantitated by ELISA. Statistics were performed by one-way ANOVA and Bonferroni post hoc analysis. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 compared to controls (None) unless otherwise indicated by brackets. ND, not significantly different compared to controls. For NO2/OVA sensitized and challenged mice, n = 6. For Th17 and Th2 adoptively transferred mice, samples from n = 3 mice were pooled prior to in vitro culturing, which was performed in triplicate. Adoptive transfer data are representative of 2 independent studies.