IM To establish a nonradioactive assay for 2′5′ oligoadenylate synthetase (25 AS) and to measure the 25AS in peripheral blood mononuclear cell (PBMC) extracts of patients with chronic hepatitis C before IFNα injec...IM To establish a nonradioactive assay for 2′5′ oligoadenylate synthetase (25 AS) and to measure the 25AS in peripheral blood mononuclear cell (PBMC) extracts of patients with chronic hepatitis C before IFNα injection, 24 hours and one month after the first injection.METHODS 25AS in cell extracts of PBMCs from 10 normal persons and 15 chronic hepatitis C patients were determined with PEI cellulose thinlayer chromatography.RESULTS The assay of 25AS in human PBMC was found to be rapid, sensitive, specific and reliable. The 25AS activity of PBMC in normal persons was in a quite low level (20%), and it was increased about tenfolds after stimulation of IFN (197%), (P<001). In 15 chronic hepatitic C patients, the basal levels of 25AS before IFN treatment were higher than those of normal persons, being much higher in the group showing poor response to IFN treatment, but 24h after the first injection of IFNα the 25AS level showed a more rapid and much greater rise in those patients with a good response.CONCLUSION 25AS may be a useful parameter of biological response during the IFN therapy..展开更多
AIM： TO examine whether 2＇-5＇oligoadenylate synthetase （OAS） gene promoter can be specifically activated by hepatitis C virus （HCV）-core protein. METHODS： Human embryo hepatic cell line L02 was transfected wit...AIM： TO examine whether 2＇-5＇oligoadenylate synthetase （OAS） gene promoter can be specifically activated by hepatitis C virus （HCV）-core protein. METHODS： Human embryo hepatic cell line L02 was transfected with pcDNA3.1-core plasmid and selected by G418. Expression of HCV-core was detected by reverse transcription polymerase chain reaction and Western blotting. The OAS promoter sequence was amplified from the genomic DNA and inserted into pGL3-basic vector. The resultant pGL3-OAS-Luci plasmid was transiently transfected into L02/core cells and luciferase activity was assayed. I^ESULTS： L02/core cell line stably expressing HCV- core protein was established. The pGL3-OAS-Luci construct exhibited significant transcriptional activity in the L02/core cells but not in the L02 cells. CONCLUSION： HCV-core protein activates the OAS gene promoter specifically and effectively. Utilization of OAS gene promoter would be an ideal strategy for developing HCV-specific gene therapy.展开更多
文摘IM To establish a nonradioactive assay for 2′5′ oligoadenylate synthetase (25 AS) and to measure the 25AS in peripheral blood mononuclear cell (PBMC) extracts of patients with chronic hepatitis C before IFNα injection, 24 hours and one month after the first injection.METHODS 25AS in cell extracts of PBMCs from 10 normal persons and 15 chronic hepatitis C patients were determined with PEI cellulose thinlayer chromatography.RESULTS The assay of 25AS in human PBMC was found to be rapid, sensitive, specific and reliable. The 25AS activity of PBMC in normal persons was in a quite low level (20%), and it was increased about tenfolds after stimulation of IFN (197%), (P<001). In 15 chronic hepatitic C patients, the basal levels of 25AS before IFN treatment were higher than those of normal persons, being much higher in the group showing poor response to IFN treatment, but 24h after the first injection of IFNα the 25AS level showed a more rapid and much greater rise in those patients with a good response.CONCLUSION 25AS may be a useful parameter of biological response during the IFN therapy..
基金Supported by National Natural Science Foundation of China,No.30671846
文摘AIM： TO examine whether 2＇-5＇oligoadenylate synthetase （OAS） gene promoter can be specifically activated by hepatitis C virus （HCV）-core protein. METHODS： Human embryo hepatic cell line L02 was transfected with pcDNA3.1-core plasmid and selected by G418. Expression of HCV-core was detected by reverse transcription polymerase chain reaction and Western blotting. The OAS promoter sequence was amplified from the genomic DNA and inserted into pGL3-basic vector. The resultant pGL3-OAS-Luci plasmid was transiently transfected into L02/core cells and luciferase activity was assayed. I^ESULTS： L02/core cell line stably expressing HCV- core protein was established. The pGL3-OAS-Luci construct exhibited significant transcriptional activity in the L02/core cells but not in the L02 cells. CONCLUSION： HCV-core protein activates the OAS gene promoter specifically and effectively. Utilization of OAS gene promoter would be an ideal strategy for developing HCV-specific gene therapy.
