LPS-induced ROS generation and changes in glutathione level and their relation to the maturation of human monocyte-derived dendritic cells.
Lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) generation and the concomitant decline in the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) were demonstrated in human monocyte-derived dendritic cells (DC). Further, their relation to the maturation of DC, characterized by the production of cytokines, up-regulation of cell surface molecules and allo-stimulatory capacity, was examined. The LPS-induced ROS generation was demonstrated using electron paramagnetic resonance spectroscopy in intact cells, and was also confirmed using laser scanning confocal microscopy. The GSH/GSSG was assesed using a glutathione assay kit. When the DC were treated with alpha-phenyl-tert-butylnitrone, the ROS generation was attenuated, but the declined GSH/GSSG was not attenuated, and only cytokine production was suppressed among the above-mentioned maturation characteristics. When the DC were treated with glutathione monoethyl ester, both the ROS generation and the declined GSH/GSSG were attenuated, and the maturation characteristics were all suppressed. These findings suggest that the LPS-induced ROS generation and the concomitant decline in GSH/GSSG occur in human monocyte-derived DC and that the former is involved in cytokine production, while the latter is involved in the up-regulation of cell surface molecules and allo-stimulatory capacity. Since the cytokine production and the allo-stimulatory capacity of DC play an important role in inflammatory and immune responses, differential regulation of the ROS generation and the declined GSH/GSSG may be useful as therapeutic tools in diseases where both responses become entangled, such as sepsis and graft-versus-host disease.
Yamada
Hiroko
H
Department of Hematology and Oncology, Kyoto University Hospital, Kyoto 606-8507, Japan.
Arai
Toshiyuki
T
Endo
Nobuyuki
N
Yamashita
Kouhei
K
Fukuda
Kazuhiko
K
Sasada
Masataka
M
Uchiyama
Takashi
T
D000954:Antigens, Surface / Q000378:metabolism; D002478:Cells, Cultured; D003497:Cyclic N-Oxides; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D005434:Flow Cytometry; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D007202:Indicators and Reagents; D018664:Interleukin-12 / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000494:pharmacology*; D007959:Lymphocyte Culture Test, Mixed; D018613:Microscopy, Confocal; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis; D009589:Nitrogen Oxides / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D015854:Up-Regulation / Q000502:physiology
D000954:Antigens, Surface; D003497:Cyclic N-Oxides; D016207:Cytokines; D007202:Indicators and Reagents; D008070:Lipopolysaccharides; D009589:Nitrogen Oxides; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12; C029217:phenyl-N-tert-butylnitrone; C042431:S-ethyl glutathione; D052247:Nitric Oxide Synthase Type II; D005978:Glutathione