Effect of AAT on oxygen consumption, lactate production, mitochondrial superoxide production, and membrane potential. (A) Ex vivo OCR in NK1.1⁺, CD11c⁺, CD4⁺CD25⁺FoxP3⁺, CD8⁺, and CD4⁺ cells from donors treated with AAT or with albumin (Control). Cells were purified from donor spleens, plated in 24-well plates and exposed to oligomycin, FCCP, and ROT plus AA (see “Materials and methods”). OCR was measured as pmol O₂/min/µg. (B) Lactate generation as measured by proton production (ECAR) in CD4⁺, CD8⁺, NK1.1⁺, CD4⁺CD25⁺FoxP3⁺, and CD11c⁺ cells from in wild-type C57/BL6 donors treated with albumin (control) or AAT (n = 5 mice per arm, experiments were done in triplicates from pooled tissue extracts). (C) Staining for mitochondrial superoxide in CD3⁺, CD4⁺, CD8⁺, NK1.1⁺, CD4⁺CD25⁺FoxP3⁺, and CD11c⁺ DCs (measured by MITOSOX). Bar plots represent the mean ± SD of 5 experiments; P values as displayed. (D) Mitochondrial TMTR in CD3⁺, CD4⁺, CD8⁺, NK1.1⁺, CD4⁺CD25⁺FoxP3⁺ and CD11c⁺ DCs. Bar plots represent the mean ± SD of 5 experiments (experiments were done in triplicates from pooled tissue extracts). P values as displayed, results were compared using the Student t test.