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Establishment of a biological assay system for human retroviral protease activity

机译:人逆转录病毒蛋白酶活性生物学检测系统的建立

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In order to obtain indicator cell lines that are exquisitely susceptible to human T-lymphotropic virus type 1 (HTLV-1), luciferase gene driven by HTLV-1 long terminal repeat (LTR) was transfected into lymphocytic H9 cells with neo gene, and cell lines were selected by G418. A cell line (H9/K301uc) was found to produce an extremely high level of luciferase only when co-cultured with HTLV-1 producer NIT-2 cells. Both in the absence and presence of a reverse transcriptase (RT) inhibitor azidothymidine, H9/K30luc cells generated similarly high luciferase activity upon co-cultivation with MT-2 cells. To develop an equivalent system for human immunodeficiency virus type 1 (HIV-1), H9/NL432 cells, which are stably infected with HIV-1 and producing a low level of the virus-like MT-2 cells for HTLV-1, were generated. Together with the indicator cell line H9/H1 luc for HIV-1 already reported, antiviral effects of some agents on HTLV-1 and HIV-1 could be readily and sensitively evaluated by similar methods. In fact, by using our system, an HIV-1 protease inhibitor, saquinavir, was demonstrated to be highly effective against HIV-1 but not against HTLV-1. (c) 2005 Elsevier SAS. All rights reserved.
机译:为了获得对人T淋巴病毒1型(HTLV-1)非常敏感的指示细胞系,将由HTLV-1长末端重复序列(LTR)驱动的萤光素酶基因与neo基因一起转染到淋巴细胞H9细胞中,通过G418选择了品系。发现仅与HTLV-1生产者NIT-2细胞共培养时,细胞系(H9 / K301uc)会产生极高水平的萤光素酶。在不存在和存在逆转录酶(RT)抑制剂叠氮胸苷的情况下,H9 / K30luc细胞在与MT-2细胞共培养时均产生相似的高荧光素酶活性。为了开发人类免疫缺陷病毒1型(HIV-1)的等效系统,分别稳定感染HIV-1并产生低水平的HTLV-1病毒样MT-2细胞的H9 / NL432细胞产生。连同已经报道的HIV-1指示细胞系H9 / H1 luc一起,可以通过类似方法轻松,灵敏地评估某些药物对HTLV-1和HIV-1的抗病毒作用。实际上,通过使用我们的系统,已证明HIV-1蛋白酶抑制剂沙奎那韦对HIV-1非常有效,但对HTLV-1无效。 (c)2005 Elsevier SAS。版权所有。

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