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dc.contributor.author이화정-
dc.date.accessioned2016-08-28T12:08:21Z-
dc.date.available2016-08-28T12:08:21Z-
dc.date.issued2009-
dc.identifier.issn0031-7012-
dc.identifier.otherOAK-5899-
dc.identifier.urihttps://dspace.ewha.ac.kr/handle/2015.oak/220244-
dc.description.abstractAlthough antiviral assays have been the most widely available biological assays for interferons (IFNs), they are less sensitive and provide considerable interassay variation. In this study, we demonstrate a new reporter cell line, which is based on HeLa cells transfected with a plasmid containing a human Mx2 promoter driving a luciferase (Luc) cDNA. To characterize the specific gene expression profiles induced by interferon alpha, we analyzed the microarray results of interferon response gene expression induced by IFN-α2a or IFN-α2b treatment with HeLa cells. We found that the Mx2 gene increased the most by treatment with both IFN-α2a and IFN-α2b. Based on this result, we designed a reporter cell line, HeLa-Mx2, suitable for determination of IFN-α. HeLa cells were stably transfected with the luciferase gene under the control of Mx2 promoter. The expression of luciferase can be easily measured for luminescence using a 96-well luminometer and has been correlated with the concentration of added IFN and cell density. In the validation results, our reporter cell line had specificity for type I IFN, but the significant effects of a number of other cytokines such as tumor necrosis factor-α, interleukin (IL)-1β, IL-2, IL-5, IL-6 and GM-CSF, or type II interferon (IFN-γ) were not observed. Moreover, the robustness of our cell line is demonstrated by the lack of an effect of the HeLa-Mx2 cell culture's age on the performance of the reporter gene assay. The reporter gene assay demonstrated reproducible dose-response curves for IFN-α2a in the range of 1-10,000 IU/ml. The 95% confidential limit and total coefficient of variation estimates ranged between 96 and 116 and 10.51% in the reducible range mentioned above, respectively. In conclusion, we established a stable IFN-responsible HeLa-Mx2 cell line, which has advantages with regard to simplicity, selectivity, and reliability over conventional cytopathic effect reduction assays used to quantify IFN-α activity. © 2009 S. Karger AG, Basel.-
dc.languageEnglish-
dc.titleValidation of a HeLa Mx2/Luc reporter cell line for the quantification of human type i interferons-
dc.typeArticle-
dc.relation.issue3-
dc.relation.volume84-
dc.relation.indexSCIE-
dc.relation.indexSCOPUS-
dc.relation.startpage135-
dc.relation.lastpage144-
dc.relation.journaltitlePharmacology-
dc.identifier.doi10.1159/000235158-
dc.identifier.wosidWOS:000270064300002-
dc.identifier.scopusid2-s2.0-68549096262-
dc.author.googleSeo Y.-J.-
dc.author.googleKim G.-H.-
dc.author.googleKwak H.-J.-
dc.author.googleNam J.-S.-
dc.author.googleLee H.J.-
dc.author.googleSuh S.-K.-
dc.author.googleBaek K.-M.-
dc.author.googleSohn Y.-W.-
dc.author.googleHong S.-H.-
dc.contributor.scopusid이화정(35069834100)-
dc.date.modifydate20170601134554-
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약학대학 > 약학과 > Journal papers
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