Fig. 1

Exposure to MCP230 induces immune responses in the lung in Th17 dependent manner. (a) Schematic of MCP230 exposure protocol. Mice were exposed to MCP230 (50 μg) by oropharyngeal aspiration on day 0 and BAL fluid (BALF) or whole-lungs were collected on day 5 post-exposure. (b) T cell subsets were quantified using flow cytometry. Pulmonary T helper cell responses were measured after vehicle or MCP230 exposure in WT and IL23p19^−/− mice. PMA/ionomycin-stimulated lung cells were stained with surface (CD3, CD4, and CD8) and intracellular (IFNγ, IL4, and IL17A) antibodies for Th1, Th2, and Th17 cells. (c) Representative pseudo color dot plots of CD4⁺ T cells gated for IFNγ, and IL17A in WT and IL23p19^−/− mice. (d) The expression of Il17a relative to Gapdh in whole-lung homogenates of WT and IL23p19^−/− mice exposed to vehicle or MCP230 was determined using RT-qPCR. e Differential cell count of BALF from vehicle or MCP230-exposed WT or IL23p19^−/− mice. f Representative photomicrographs of BALF cells collected from WT and IL23p19^−/− mice exposed to vehicle or MCP230. Arrowheads represent the neutrophils (black) and lymphocytes (white). Data represent mean ± SEM from 3 to 5 mice. ^ap < 0.05, compared to WT Vehicle group. ^bp < 0.05, compared to WT MCP230 group, one-way ANOVA with Tukey’s multiple comparisons test