Joh T, Fujita M, Tanaka Y, Shiku H
Department of Oncology, Nagasaki University School of Medicine, Japan.
Gene. 1995 Aug 19;161(2):227-30. doi: 10.1016/0378-1119(95)00249-6.
We prepared the murine myeloma cell line NS-1, which stably expressed the human T lymphotropic virus type I (HTLV-I) env gene. The plasmid BCMGEnv was constructed from the episomal vector BCMGSNeo, which was primarily derived from bovine papilloma virus. Transfected env expression was detected by Northern blotting, as well as by flow cytometry using envelope protein-specific monoclonal antibodies (mAb). Expression was detectable for at least seven months. The env transfectants induced syncytium formation which is characteristic of HTLV-I-infected cells, in the human uterine cervical cancer line, HeLa, and the rat cell line, XC. The requirement of envelope proteins for syncytium formation was confirmed by an inhibition assay with envelope protein-specific mAb. Therefore, env transfectants are not only stable, but also have its specific biological function. This system may be useful to analyze the initial steps of viral attachment to the cell surface and to search for the HTLV-I receptor.
我们制备了稳定表达人嗜T淋巴细胞病毒I型(HTLV-I)env基因的小鼠骨髓瘤细胞系NS-1。质粒BCMGEnv由游离载体BCMGSNeo构建而成,该载体主要来源于牛乳头瘤病毒。通过Northern印迹法以及使用包膜蛋白特异性单克隆抗体(mAb)的流式细胞术检测转染后的env表达。至少七个月内均可检测到表达。env转染子在人子宫颈癌细胞系HeLa和大鼠细胞系XC中诱导了合胞体形成,这是HTLV-I感染细胞的特征。通过包膜蛋白特异性mAb的抑制试验证实了包膜蛋白对合胞体形成的必要性。因此,env转染子不仅稳定,而且具有其特定的生物学功能。该系统可能有助于分析病毒附着于细胞表面的初始步骤以及寻找HTLV-I受体。