The phagocytosis-associated respiratory burst in human monocytes is associated with increased uptake of glutathione.
During the phagocytic respiratory burst, oxygen is converted to potent cytotoxic oxidants. Monocytes and macrophages are potentially long-lived, and we have hypothesized that protective mechanisms against oxidant stress are varied and fully expressed in these cells. We report here that the respiratory burst in monocytes is accompanied by an increase in the uptake of [35S]glutathione ([35S]GSH) after 20-30 min to levels up to 10-fold greater than those at baseline. By 30 min, 49% of the cell-associated radioactivity was in the cytosol, 41% was in membrane, and 10% was associated with the nuclear fraction. GSH uptake was inhibited by catalase, which removes hydrogen peroxide (H2O2), and micromolar H2O2 stimulated GSH uptake effectively in monocytes and also lymphocytes. Oxidation of GSH to glutathione disulfide with H2O2 and glutathione peroxidase prevented uptake. Acivicin, which inhibits GSH breakdown by gamma-glutamyl transpeptidase (GGT), had no effect on the enhanced uptake seen during the respiratory burst. Uptake of cysteine or cystine, possible products of GGT activity, stayed the same or decreased during the respiratory burst. These results suggest that a GGT-independent mechanism is responsible for the enhanced GSH uptake seen during the respiratory burst. We describe here a sodium-independent, methionine-inhibitable transport system with a Km (8.5 microM) for GSH approximating the plasma GSH concentration. These results suggest that monocytes have a specific GSH transporter that is triggered by the release of H2O2 during the respiratory burst and that induces the uptake of GSH into the cell. Such a mechanism has the potential to protect the phagocyte against oxidant damage.
Seres
T
T
Department of Pediatrics, Yale University School of Medicine, New Haven, CT 06520, USA.
Knickelbein
R G
RG
Warshaw
J B
JB
Johnston
R B
RB
D003545:Cysteine / Q000037:antagonists & inhibitors / Q000378:metabolism; D003553:Cystine / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism; D008262:Macrophage Activation; D009000:Monocytes / Q000276:immunology / Q000378:metabolism*; D018384:Oxidative Stress / Q000276:immunology; D010587:Phagocytosis / Q000276:immunology*; D016897:Respiratory Burst / Q000276:immunology*; D012964:Sodium / Q000502:physiology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D003553:Cystine; D012964:Sodium; D006861:Hydrogen Peroxide; D005978:Glutathione; D003545:Cysteine; D013755:Tetradecanoylphorbol Acetate