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

Figure 2

The persistent presence of fungi in lung injury induced by Aspergillus fumigatus is related to Tregs. (a) Increased CD4⁺CD25⁺Foxp3⁺ Treg proportion significant in PBMCs of patients infected with A. fumigatus (). (b) Foxp3 mRNA levels in the lungs were measured with q-PCR. Relative expression levels of the genes were expressed with the GAPDH housekeeping gene as an internal reference (/group). The expression of Foxp3 protein levels in the lungs was measured with Western blotting. Relative expression levels were expressed with the GAPDH as an internal reference (/group). (c) CD4⁺CD25⁺ T cells and CD4⁺CD25⁺Foxp3⁺ Tregs in the spleen were detected by flowcytometry, and FlowJo10 analyzed the proportions as prior described in Section 2. Increased CD4⁺CD25⁺ T cell and CD4⁺CD25⁺Foxp3⁺ Treg proportion significantly in the spleen of A. fumigatus infection mice (/group).Decreased susceptibility of mice to A. fumigatus infection after Treg depletion. (d) After pretreatment with CD25-neutralizing antibodies, the number of Treg cells in the spleen of mice with A. fumigatus infection group was significantly reduced, each group (/group). (e) Fungal load (colony-forming unit) after 1 day of infection (/group). Experiments were done at least three times. Data are presented as deviation. Statistically significant difference () against noninfected control. (f) Lungs from each experimental group were processed for histological examination after H&E staining. Lung injury scores were evaluated by the method described previously. , , , and by Student’s unpaired two-tailed test and the one-way ANOVA followed by the Tukey post hoc test. Error bars represent SEM.

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