Figure 6.

Metformin Rejuvenates OXPHOS and Reduces Inflammatory Cytokine Production in TB10.4-TET⁺ Cells

(A) Experimental timeline.

(B and C) CFUs in the lungs of mice treated with or without metformin (MET) over the course of infection (B) and corresponding pathology scores performed by morphometric analysis (Visiopharm) on lung tissue sections stained with H&E (C). Total CD8⁺ T cells purified from the lungs of uninfected (UI) or Mtb-infected mice treated with or without MET were analyzed using XF or flow cytometry.

(D) OCR at baseline.

(E–K) OCR measured during a cell mito stress test (CMST) (E) with associated bioenergetic parameters (F) and corresponding phenogram (G) after uncoupling with FCCP at D21 post-infection and (H–K) D35 post-infection.

(L) Glycolysis, measured by the change in ECAR pre- and post-injection with glucose.

(M) Frequency of Mtb-specific, TB10.4-TET⁺ cells in lungs and spleen after MET.

(N) Cytokine production in TB10.4-TET⁺ cells after re-stimulation with PMA/ionomycin.

(O) Polyfunctionality on the basis of co-expression of IFNγ, TNF-α, and IL-2 (single, double, or triple combinations) of TB10.4-TET⁺ cells in (N).

A&R, antimycin A and rotenone; F, carbonyl cyanide-4-[trifluoromethoxy] phenylhydrazone or FCCP; O, oligomycin. ^∗ECAR values were normalized to a zero baseline. Statistics, unless otherwise indicated, are relative to UI. Data are representative of two independent experiments (N = 5 mice per group). “p ≤ 0.05, ^#p ≤ 0.01, ^∞p ≤ 0.005, and ^∗p ≤ 0.001 by one-way ANOVA or unpaired Student’s t test. Error bars are mean ± SD.