1. Dysfunctional antifungal neutrophils have impaired expression of IFN-inducible genes.
Dysfunctional antifungal neutrophils have impaired expression of IFN-inducible genes. (A) ROS production by airway-infiltrating neutrophils isolated from CCR2-depleted mice (red histogram) or control littermates (gray histogram) 48 hours after infection. Graph shows mean ± SEM of ROS mean fluorescence intensity (MFI) for five mice per group assayed by FACS. FITC, fluorescein isothiocyanate. (B) Af pulmonary fungal burden 48 hours after infection. CFU, colony-forming units. (C) Differential gene expression as assessed by RNA-seq of pulmonary neutrophils isolated from uninfected controls (naïve) or mice infected with Af for 48 hours. Heat map depicts the top 100 genes expressed at a fold change of >2.5 in control neutrophils more than in neutrophils from CCR2-depleted mice. (D) Venn diagram of differentially expressed genes. Predicted upstream regulators of the 231 genes as determined with Ingenuity Pathway Analysis (IPA) software. (E to J) Kinetics of IFN expression in the lung of mice at different times after Af infection. (E to G) Gene expression as determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) using TaqMan probes. Data are means ± SEM of five mice per time point. Graphs of (H) to (J) show mean ± SEM of each cytokine as measured by ELISA in five mice per time point. (K to M) Pulmonary qRT-PCR of IFN gene expression at 3 hours (K) or 48 hours (L and M) after Af infection. *P < 0.05; **P < 0.01; ***P < 0.001, calculated by Mann-Whitney nonparametric test or Kruskal-Wallis test using Prism software.
Vanessa Espinosa et al. Sci. Immunol. 2017;2:eaan5357
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