Regardless of adjuvant, a combined eosinophilic/neutrophilic inflammatory profile was observed in all OVA sensitized groups following OVA challenge. Open in a separate window Figure 1 Serum OVA-specific IgE and airway inflammatory reactions are enhanced in OVA/CFA sensitized mice. cellular GSK1059865 sources of IL-17 were assessed from bronchoalveolar lavage and/or lungs. The part of T cells in these reactions was tackled in OVA/CFA sensitized mice using a T cell antibody. Results Following OVA challenge, all mice exhibited combined eosinophilic/neutrophilic airway inflammatory profiles and elevated serum OVA-IgE. Whereas OVA/alum sensitized mice experienced moderate swelling and AHR, OVA/CFA sensitized mice experienced significantly higher swelling but lacked AHR. This correlated with a shift in IL-17 production from CD4+ to T cells. Additionally, OVA/CFA sensitized mice, given a TCR stimulatory antibody, showed improved frequencies of IL-17- T cells and diminished airway reactivity and eosinophilia. GSK1059865 Conclusions Therefore, the conditions of antigen sensitization influence the profile of cells that create IL-17, the balance of which may then modulate the airway inflammatory reactions, including AHR. The possibility for IL-17- T cells to reduce AHR and powerful eosinophilic swelling provides evidence that therapeutic methods focused on stimulating and increasing airway IL-17- T cells may be an effective alternate in treating steroid resistant, severe asthma. Electronic supplementary material The online version of this article (doi:10.1186/s12931-014-0090-5) contains supplementary material, which is available to authorized users. gene manifestation before quantification from the comparative threshold cycle method to obtain the gene manifestation levels from lungs of OVA sensitized and challenged mouse organizations, relative to the saline control group [30]. Quantitative analysis of BAL fluid mediators BAL fluid cytokine and chemokine levels were quantified with the Q-View Imager using the 16-plex mouse cytokine display (Quansys Biosciences, Logan, Utah, USA). IL-13 levels in the BAL fluid were quantified using the ELISA Ready-SET-Go kit (ebioscience, San Diego, California, USA). Statistical analysis Data are indicated as the mean?+SEM. Multiple comparisons (we.e. antigen- and adjuvant-dependent effects) were analyzed by two-way ANOVA, followed by the Holm-Sidak post hoc test. Single comparisons (between the 3 OVA-sensitized organizations) were analyzed by one-way ANOVA, followed by the Holm-Sidak post hoc test. Single comparisons (between the 2 antibody treated organizations) were analyzed by an unpaired, two-tailed t-test. p-values less than 0.05 were considered statistically significant. Numbers and statistics were analyzed using GraphPad Prism 6 (La Jolla, California, USA). Results Enhanced airway swelling, but lack of AHR, in mice sensitized to OVA in the presence of CFA In order to establish a mixed model of sensitive asthma in which the IL-17 response could be assessed within an Th2 environment, we intraperitoneally (IP) sensitized mice with OVA in the absence (OVA/sal group) or presence of the adjuvants, alum (OVA/alum group) or CFA (OVA/CFA group). We confirmed induction of several classic features associated with sensitive airways disease and differentiated OVA-specific () from adjuvant-specific (*) effects (Number?1). OVA-IgE was selectively recognized in all OVA sensitized and challenged mice and was present at significantly higher levels in OVA/CFA mice (Number?1A). Total cells, eosinophils, neutrophils and lymphocytes were significantly improved in the BAL fluid of OVA/CFA sensitized mice (solid pub) compared to the CFA control (striped pub). Cdkn1a In contrast, inflammation was not significantly changed in OVA/sal mice and only eosinophils were significantly improved in OVA/alum sensitized mice (Number?1B). Moreover, following OVA challenge, OVA/CFA mice experienced significantly more GSK1059865 macrophages, eosinophils, neutrophils and lymphocytes, resulting in 3 and 5.5 fold more total cells recovered compared to OVA/alum and OVA/sal mice, respectively. With regard to BAL fluid cell frequencies, eosinophils were increased in all OVA-sensitized mice compared to their respective GSK1059865 controls, primarily at the expense of macrophages (Additional file 1: Number S1). OVA/alum sensitized and challenged mice experienced higher frequencies of BAL fluid eosinophils than OVA/sal mice, while OVA/CFA mice experienced greater frequencies.