An Elucidation of Neutrophil Functions against Mycobacterium tuberculosis Infection
Figure 3
GSH measurements in neutrophils infected with H37Rv and treated with NAC and L-GSH ( ). GSH levels were measured in isolated neutrophils from healthy subjects that were infected in vitro with H37Rv and treated with GSH-enhancing agents such as NAC and L-GSH, by spectrophotometry using an assay kit. Briefly, H37Rv-infected neutrophils (3 × 105) were lysed at 24 hours after infection with ice cold 5% 5-sulfosalicylic acid dehydrate solution. Supernatants collected after centrifugation were analyzed for total GSH ((a)) and GSSG (c) as per manufacturer’s instructions. rGSH ((b)) was calculated by subtracting measured GSSG concentrations from the measured total GSH concentrations. All GSH measurements were normalized with total protein levels. Proteins in the cell lysates of neutrophils were measured by Bradford’s method using a Coomassie protein assay reagent. (d) illustrates ratio of rGSH to GSSG expressed as percentage of total GSH in neutrophil lysates. Graph shows mean ± standard error.
