Aspergillus fumigatus Influences Gasdermin-D-Dependent Pyroptosis of the Lung via Regulating Toll-Like Receptor 2-Mediated Regulatory T Cell Differentiation

Figure 4

TLR2 affects the proliferation of CD4⁺CD25⁺Foxp3⁺ Treg in lung injury caused by Aspergillus fumigatus. (a) CD4⁺CD25⁺ T cells and CD4⁺CD25⁺Foxp3⁺ Tregs in the spleen were detected by flowcytometry, and the proportions were analyzed by FlowJo10 as prior described in Section 2. Reduced CD4⁺CD25⁺ T cell and CD4⁺CD25⁺Foxp3⁺ Treg proportion significantly in the spleen of A. fumigatus infection mice (/group). (b) Foxp3 mRNA levels in the lungs were measured with qRT-PCR (/group). (c) The expression of Foxp3 protein in lungs of C57BL/6 mice and mice was infected with A. fumigatus for 3 days which was detected by Western blot and analyzed by imager systems described in Section 2 (/group). CD4⁺ T cells were sorted from the spleens of wild-type C57BL/6 mice and cultured. Tregs were detected by flow cytometry on day 3 and analyzed by FlowJo10. (d) CD4⁺ T lymphocytes differentiated fewer CD4⁺CD25⁺ T cells and CD4⁺CD25⁺Foxp3⁺ Tregs after treatment with C29. In each group, ; three replicate experiments were performed three times. ns: not significant; , , , and by the one-way ANOVA and the two-way ANOVA followed by the Tukey post hoc test. Error bars represent SEM.

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