Tajbakhsh S, Kiss G, Lee P E, Seligy V L
Division of Biological Sciences, National Research Council of Canada, Ottawa, Ontario.
Virology. 1990 Jan;174(1):264-75. doi: 10.1016/0042-6822(90)90074-2.
Comparative studies were carried out using two different insect cell lines, Aedes albopictus and Estigmene acrea, for Tipula iridescent virus (TIV) propagation. Light microscope autoradiography showed viral DNA present in viroplasmic centers (VCs) and an inhibition of nuclear DNA synthesis. These VCs appeared to be morphologically similar in both cell lines when examined by light and electron microscopy. Radiolabeled cDNA was synthesized from RNA samples obtained from infected cells at different times after infection and hybridized to TIV DNA digested with various restriction endonucleases. The results indicated that the pattern of transcription and the kinetics of TIV infection were qualitatively similar in both cell lines. The major TIV DNA components, L (greater than 174 kbp) and S1 (10.8 kbp) that are found in virions in approximately equivalent amounts, were made in both infected cell lines. However, the infected cell lines produced S1 DNA at higher levels relative to L than in virions. The cDNA hybridization studies also revealed that the S1 DNA has sequences that are transcribed and are TIV specific. While VC morphology, levels of L and S1 DNA synthesis, transcription, and capsid protein synthesis were similar in both cell lines, time course electron microscope studies revealed that progeny virions were detected only in the VCs of E. acrea cells and not in the VCs of A. albopictus cells, even by 96 hr p.i. These data suggest that the A. albopictus C6/36 cell line is semipermissive for TIV replication.
使用两种不同的昆虫细胞系,即白纹伊蚊和草地贪夜蛾,对大蚊虹彩病毒(TIV)进行增殖的比较研究。光学显微镜放射自显影显示病毒DNA存在于病毒质中心(VCs),并且抑制了核DNA合成。当通过光学显微镜和电子显微镜检查时,这两种细胞系中的这些VCs在形态上似乎相似。从感染后不同时间的感染细胞获得的RNA样本中合成放射性标记的cDNA,并与用各种限制性内切酶消化的TIV DNA杂交。结果表明,两种细胞系中TIV的转录模式和感染动力学在质量上相似。在病毒粒子中以大致等量存在的主要TIV DNA成分L(大于174 kbp)和S1(10.8 kbp),在两种感染的细胞系中均有产生。然而,与病毒粒子相比,感染的细胞系产生的S1 DNA相对于L的水平更高。cDNA杂交研究还表明,S1 DNA具有被转录且是TIV特异性的序列。虽然两种细胞系中的VC形态、L和S1 DNA合成水平、转录和衣壳蛋白合成相似,但时间进程电子显微镜研究表明,即使在感染后96小时,也仅在草地贪夜蛾细胞的VCs中检测到子代病毒粒子,而在白纹伊蚊细胞的VCs中未检测到。这些数据表明,白纹伊蚊C6/36细胞系对TIV复制是半允许的。