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一种用于人嗜T淋巴细胞病毒1型(HTLV-1)感染性和药物发现的新型中通量生物学检测系统。

A novel medium-throughput biological assay system for HTLV-1 infectivity and drug discovery.

作者信息

Abdizadeh Mojtaba Fattahi, Makvandi Manoochehr, Samarbafzadeh Alireza, Azadmanesh Kayhan

机构信息

Department of Microbiology, Faculty of Medicine, Sabzevar University of Medical Sciences, Sabzevar, Iran.

Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

出版信息

Iran J Basic Med Sci. 2017 Oct;20(10):1109-1118. doi: 10.22038/IJBMS.2017.9417.

Abstract

OBJECTIVES

Here, a reporter cell line containing two reporter vectors were developed, in order to monitor the Human T-Lymphotropic Virus type1(HTLV-1) infectivity and the cell viability simultaneously.

MATERIALS AND METHODS

The reporter cell line was constructed by stably transfected baby hamster's kidney cell line (BHK-21), with the genomes expressing two different reporters in separate plasmids. The first reporter gene is transactivated by the HTLV-1 tax protein, while the second reporter is continuously expressed when introduced into a mammalian cell. In order to show its functionality, the effect of the drug mix on HTLV-1 was assayed by this system and was compared to the results obtained by other methods.

RESULTS

HTLV-1 reporter cell line was found to produce high level of luciferase when co-cultured with MT-2 and Hut-102 cells but not with Jurkat cell. Moreover, the combination therapy against HTLV-1 can reduce luciferase expression of the cell when co-cultured with MT-2 and Hut-102 comparable to the ELISA (R=0.932, -value =0.002). In addition, the results revealed the superiority of the present system over the molecular methods.

CONCLUSION

The results demonstrated that the biological assay system is a beneficial tool for the medium-throughput anti-HTLV-1 drug screening and inhibitory effect.

摘要

目的

在此,开发了一种包含两个报告载体的报告细胞系,以便同时监测1型人类嗜T淋巴细胞病毒(HTLV-1)的感染性和细胞活力。

材料与方法

通过将表达两种不同报告基因的基因组分别稳定转染到幼仓鼠肾细胞系(BHK-21)中构建报告细胞系。第一个报告基因由HTLV-1 tax蛋白反式激活,而第二个报告基因在导入哺乳动物细胞时持续表达。为了展示其功能,用该系统检测了药物组合对HTLV-1的作用,并与其他方法获得的结果进行了比较。

结果

发现HTLV-1报告细胞系与MT-2和Hut-102细胞共培养时可产生高水平的荧光素酶,但与Jurkat细胞共培养时则不然。此外,与MT-2和Hut-102共培养时,针对HTLV-1的联合疗法可降低细胞的荧光素酶表达,与酶联免疫吸附测定法相当(R=0.932,P值=0.002)。此外,结果显示本系统优于分子方法。

结论

结果表明,该生物学检测系统是用于中通量抗HTLV-1药物筛选和抑制作用的有益工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b895/5673695/31dbd80db7ae/IJBMS-20-1109-g001.jpg

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