Autophagy induced by Mtb-Ag in neutrophils from HR and LR TB patients. Neutrophils were incubated at 2.5 × 106 cells/ml for 2 h. Then, autophagy levels were evaluated by (A and C) flow cytometry against intracellular LC3B, (B) immunofluorescence against LC3B. (A) Each bar represents the mean of percentage of LC3B+ neutrophils ± SEM. Black and grey bars corresponding to HR or LR patients respectively. (B) Representative immunofluorescence images of 1 HR and 1 LR TB patient are shown. (C) Representative dot plots for a HR TB patient (left panel) and a LR TB patient (right panel) are shown. Negative controls (upper panel) represent the samples stained only with the secondary antibody for the indirect method. Statistical differences were calculated using the Wilcoxon signed rank test for paired samples (***P < 0.001; ****P < 0.0001) or the Mann-Whitney nonparametric test for unpaired samples (###P < 0.001). N, HR = 13; N, LR = 14.
