FIG. 7.
Activated CD8+ T cells were analyzed for expression of iGSH using monochlorobimane (mBCl). Briefly, cells were preloaded for 15 min with 10-μM mBCl, which forms blue fluorescent adducts with iGSH. Immediately before flow cytometry analysis, propidium iodide was added. For mBCl, cells were excited with a violet (UV) 405-nm laser, and emission was acquired with a 440/40 filter. Changes in the iGSH are reflected by the appearance of populations of cells with differences in mBCl fluorescence. Histogram represents the level of fluorescence from stained samples that were gated for high or low cell surface expression of CD45RA. CD45RA is primarily expressed on naïve T cells (CD45RAhi) and is lost after TCR stimulation (CD45RAlo). Data shown are from one of three experiments with similar results. iGSH, intracellular glutathione; RA, rheumatoid arthritis.