Autophagy prevents early proinflammatory responses and neutrophil recruitment during Mycobacterium tuberculosis infection without affecting pathogen burden in macrophages
Fig 4
Autophagy suppresses neutrophil recruitment independent of mitophagy and inflammasome activation during M. tuberculosis infection.
(A) Proportion of CD45+ cells that are neutrophils (CD45+Ly6G+CD11b+) in the lung at 14 dpi in Mtb-GFP infected Becn1fl/fl (n = 5), Becn1fl/fl-LysM-Cre (n = 7), Caspase1/11-/-/Becn1fl/fl (n = 9), Caspase1/11-/-/Becn1fl/fl-LysM-Cre (n = 13), Atg5fl/fl (n = 5), Atg5fl/fl-LysM-Cre (n = 5), Caspase1/11-/-/Atg5fl/fl (n = 4), or Caspase1/11-/-/Atg5fl/fl-LysM-Cre (n = 5) mice reported as a ratio relative to the average neutrophil frequency in corresponding floxed control mice. (B) Lung burden at 14 dpi from right lobes of the lung in Mtb-GFP infected Becn1fl/fl (n = 5), Becn1fl/fl-LysM-Cre (n = 7), Caspase1/11-/-/Becn1fl/fl (n = 9), Caspase1/11-/-/Becn1fl/fl-LysM-Cre (n = 13), Atg5fl/fl (n = 5), Atg5fl/fl-LysM-Cre (n = 5), Caspase1/11-/-/Atg5fl/fl (n = 4), or Caspase1/11-/-/Atg5fl/fl-LysM-Cre (n = 5) mice. The legend in 4A should be used for 4B too. (C) Kaplan–Meier curve of survival proportions during Mtb-GFP infection of Atg5fl/fl, Atg5fl/fl-LysM-Cre, Caspase1/11-/-/Atg5fl/fl, and Caspase1/11-/-/Atg5fl/fl-LysM-Cre mice. (D) Proportion of CD45+ cells that are neutrophils (CD45+Ly6G+CD11b+) in the lung at 14 dpi in Mtb-GFP infected WT (n = 4), Parkin-/- (n = 3), or Pink1-/- (n = 3) mice reported as a ratio relative to the average neutrophil frequency in WT mice. (E) Lung burden from the right lobe of the lung at 14 dpi in Mtb-GFP infected WT (n = 4), Parkin-/- (n = 3), or Pink1-/- (n = 3) mice. Statistical differences were determined by log-rank Mantel–Cox test (C) and one-way ANOVA and Šídák multiple comparison test (A, B, D, and E). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Differences that are not statistically significant are designated as ns. Pooled data from at least 2 separate experiments is graphed where each data point is from 1 biological replicate. The individual numerical values used to generate the graphed data in Fig 4, the statistical analyses performed to analyze these data, and the p values from these statistical tests are in S4 Data. dpi, days postinfection; WT, wild-type.