Fig. 2: JHU083 administration reduces Mtb proliferation and lung pathology in mice.

a Schematic of the mouse experiments. Six to ten weeks old 129S2 or C3HeB/FeJ mice (n = 4–10/group) were aerosol infected with ~200–300 CFU of Mtb H37Rv. The mice were treated with JHU083 or RIF by oral gavage starting one day after infection. 1 mg/kg of JHU083 was given daily for the first five days, and then the dose was reduced to 0.3 mg/kg daily (5/7, M-F). 12.5 mg/kg RIF was given daily orally for 5-weeks. b Lung bacillary burden in 129S2 mice treated with JHU083 or PBS or RIF (n = 5/group for week 2 & n = 4/group for week 5). Mice were sacrificed at day 0, week 2, and week 5 post-infection/treatment. The lungs were harvested, homogenized, diluted, and plated on 7H11 selection plates. After 21–25 days, the colonies were counted, and counts were transformed into log₁₀ values and plotted. c DON levels in lungs of JHU083-treated, Mtb-infected 129S2 mice (n = 5/group) at 2 weeks post-infection, 30–40 min after receiving a 1 mg/kg dose of JHU083 (the tissue Tmax of JHU083 is 30 min post-oral dosing; full dosing details are in the Methods). DON levels were determined using LC/MS-based targeted metabolomics. d Survival of 129S2 mice treated with JHU083 or PBS. e Gross lung weight of 129S2 mice (n = 10/group) at weeks 2 and 5 post infection/treatment. f Histopathology of lungs isolated from C3HeB/FeJ mice infected with Mtb H37Rv at week 4.5 post-infection/treatment. The lungs were formalin fixed, sectioned, and stained with hematoxylin and eosin (H&E). g Quantitation of the lung granuloma areas in C3HeB/FeJ mice infected with Mtb H37Rv at week 4.5 post-infection/treatment (n = 6). h Quantitation of the lung granuloma areas in the 129S2 mice lungs infected with Mtb H37Rv at week 5 post-infection/treatment (n = 3). Both total granuloma area (GA) and lung area (LA) areas were measured using ImageScope software (Leica). The percent granuloma area (%GA) was calculated using the formula (%GA = (GA X 100)/LA). Data were plotted as mean ± SEM. Statistical significance was calculated using a two-tailed student t-test considering unequal distribution. For survival curve, log-rank (Mantel-Cox) and Gehan-Breslow-Wilcoxon tests were used and yielded similar p-value. The exact p-values are provided in the Source Data file. * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. CFU stands for colony-forming units. All the experiments were repeated at least twice. The mouse clipart shown in 2a was created using Biorender.com (Tornonto, Canada) by SP.