Host range of human T-cell leukemia virus type I analyzed by a cell fusion-dependent reporter gene activation assay
Host range of human T-cell leukemia virus type I analyzed by a cell fusion-dependent reporter gene activation assay
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DOI：
10.1006/viro.1998.9530
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发表时间：
1999-02-15
期刊：
影响因子：
3.7
通讯作者：
Matsuura, Y
中科院分区：
文献类型：
--
作者：
Okuma, K;Nakamura, M;Matsuura, Y
我们构建了一个灵敏的定量检测系统，以检查人类T细胞白血病病毒I型（HTLV-I）包膜（env）糖蛋白介导的细胞融合，其中T7 RNA聚合酶在供体细胞共表达env糖蛋白激活受体细胞的报告基因后，细胞融合。通过免疫印迹和免疫荧光分析，在用表达质粒转染的293T细胞中观察到HTLV-Ⅰ env糖蛋白（gp46和gp21）的有效表达。表达env糖蛋白的细胞也表现出自身融合。通过将供体细胞与用在T7启动子下具有荧光素酶基因的报告质粒转染的受体细胞共培养，在细胞融合时观察荧光素酶的表达。荧光素酶基因的激活被抗env中和抗体或对应于env gp21的合成肽抑制，从而表明细胞融合由供体细胞中表达的HTLV-1 env糖蛋白特异性介导。广泛的细胞系表现出对HTLV-I env介导的细胞融合的敏感性。因此，新建立的检测系统可能为研究HTLV-Ⅰ env糖蛋白介导的融合机制和鉴定HTLV-Ⅰ受体提供了一种有效的方法。(C)北京：科学出版社.
We constructed a sensitive and quantitative assay system to examine human T-cell leukemia virus type I (HTLV-I) envelope (env) glycoprotein-mediated cell fusion in which T7 RNA polymerase in donor cells coexpressing env glycoproteins activates a reporter gene in recipient cells upon cell fusion. An efficient expression of HTLV-I env glycoproteins (gp46 and gp21) was observed in 293T cells transfected with an expression plasmid by both immunoblot and immunofluorescence analyses. The cells expressing env glycoproteins also exhibited self-fusion. By cocultivating the donor cells with recipient cells transfected with a reporter plasmid possessing the luciferase gene under the T7 promoter, the expression of luciferase was observed upon cell fusion. The activation of the luciferase gene was inhibited by either anti-env neutralizing antibody or synthetic peptide corresponding to env gp21, thus indicating the cell fusion to be specifically mediated by the HTLV-I env glycoproteins expressed in the donor cells. A broad range of cell lines exhibited susceptibility to HTLV-I env-mediated cell fusion by this assay. This newly established assay system may thus provide an efficient way both to study the fusion mechanisms mediated by HTLV-I env glycoproteins and to identify the HTLV-I receptor(s). (C) 1999 Academic Press.