[Establishment of superoxide production assay system using Epstein-Barr virus transformed cell line with chemiluminescence].
We established the system to detect superoxide produced by Epstein Barr virus lymphoblastoid cell line (EB-LCL). Superoxide production of EB-LCL was evaluated by measuring chemiluminescence (CL) enhanced with addition of horseradish peroxidase (HRP). Using this system, we measured CL of EB-LCL established from 13 patients with chronic granulomatous disease (CGD) and 8 normal individuals. Significant elevation of CL was observed in all control EB-LCLs, however, no remarkable CL was seen in any patients' EB-LCLs. We examined the effect of recombinant human interferon gamma (rh-IFN-gamma) and granulocyte colony stimulating factor (G-CSF) on CL of EB-LCL in vitro. With addition of rh-IFN-gamma, CL of normal control EB-LCL was significantly enhanced (p < 0.05), on the other hand, G-CSF was shown to have no effect. No significant CL was observed in any CGD patients' EB-LCLs even with addition of rh-IFN-gamma or G-CSF. It was suggested that superoxide produced by EB-LCL detected in this system was dependent on the same NADPH oxidase system which presents in phagocyte.
Furuta
H
H
Department of Pediatrics, School of medicine, Hokkaido University.
Ariga
T
T
Sakiyama
Y
Y
Matsumoto
S
S
D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D002461:Cell Line, Transformed; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006105:Granulomatous Disease, Chronic / Q000378:metabolism / Q000473:pathology; D004854:Herpesvirus 4, Human; D006735:Horseradish Peroxidase; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008163:Luminescent Measurements; D009247:NADH, NADPH Oxidoreductases / Q000502:physiology; D019255:NADPH Oxidases; D011994:Recombinant Proteins; D013481:Superoxides / Q000378:metabolism*
D011994:Recombinant Proteins; D013481:Superoxides; D016179:Granulocyte Colony-Stimulating Factor; D007371:Interferon-gamma; D006735:Horseradish Peroxidase; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases