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Additional file 1: Supplementary figure 1. pH of media culture with lactic acid. Different concentration of lactic acid were added to RPMI 1640 medium supplemented with 10% FBS. The pH were measured with pH meter. Supplementary figure 2. The purity of T cells is represented. PBMCs were seeded into 24-well plates (1.5��106 cells/well) and cultured in RPMI1640 containing 10���S and 100IU hIL-2. To activate and enrich T cells, PBMCs were cultured with 3 ug/ml anti-CD3 antibody and 10 ug/ml anti-CD28 antibody. After 4 days of incubation at 37��C purity of T cells was assayed by flow cytometry using APC conjugated anti-human CD3. Supplementary figure 3. Viability of raji cells were treated with different concentration of DCA. 2 �� 105 Raji cells were treated with various concentrations of DCA. The apoptosis was detected by flow cytometry using the PI staining. DCA: dichloroacetate PI: propidium iodide. Supplementary figure 4. DCA reduced tumor-derived lactate. 2 �� 105 Raji cells were treated in the presence or absence of mitomycin C. Lactate concentration in the tumor cells supernatant was measure after 24h and 48h. Lactate was measured using the colorimetric assay. Data are presented as mean �� SD from a representative experiment (n = 3). Two-way ANOVA was used to examine the difference between groups. Tukey���s post hoc test was used to compare means. Supplementary table 1. Primers used for gene expression analysis through real-time PCR.

Additional file 1: Fig. S1. PCA plots for batch effect removal using ComBat-seq in Whole Blood samples: The PCA plots show the samples of RNA-seq datasets before (A) and after (B) batch effect removal. Fig. S2. PCA plots for batch effect removal using ComBat-seq in nasopharyngeal samples: The PCA plots show the samples of RNA-seq datasets before (A) and after (B) batch effect removal.

Multiple roles have been indicated for reactive oxygen species (ROS) in the immune system in recent years. ROS have been extensively studied due to their ability to damage DNA and other subcellular structures. Noticeably, they have been identified as a pivotal second messenger for T-cell receptor signaling and T-cell activation and participate in antigen cross-presentation and chemotaxis. As an agent with direct toxic effects on cells, ROS lead to the initiation of the autoimmune response. Moreover, ROS levels are regulated by antioxidant systems, which include enzymatic and nonenzymatic antioxidants. Enzymatic antioxidants include superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase. Nonenzymatic antioxidants contain vitamins C, A, and E, glutathione, and thioredoxin. Particularly, cellular antioxidant systems have important functions in maintaining the redox system homeostasis. This review will discuss the significant roles of ROS generation and antioxidant systems under normal conditions, in the immune system, and pathogenesis of multiple sclerosis.