Osman H K, Wells C, Baboonian C, Kangro H O
Department of Virology, St. Bartholomew's Hospital Medical College, West Smithfield, London, United Kingdom.
J Med Virol. 1993 Apr;39(4):303-11. doi: 10.1002/jmv.1890390409.
In order to optimize viral antigen production, the growth characteristics of human herpesvirus-6 (HHV-6) (strain AJ) were studied in two cell lines: HSB-2 and JJHAN. The cells were infected with different multiplicities of infection (moi) and viral growth was monitored by appearance of cytopathic effect (CPE), total and viable cell count, immunofluorescence test, immunoblotting, and electron microscopy. Although > or = 70% of JJHAN cells showed CPE when infected at high moi, only 5% of the cells contained viral antigens when tested with immunofluorescence. In contrast the percentage of cells showing fluorescence in HSB-2 cells reached > or = 30% when infected at > or = 1:50. More than 10 polypeptides of molecular weight ranging between 31-140 kD appeared in the HSB-2 cultures by immunoblotting while only 3 polypeptides were detected in the JJHAN cultures at high moi. Different stages of virus maturation were seen in the HSB-2 cells by electron microscopy but the replication of the virus in JJHAN cells appeared to be restricted. For the purpose of antigen production the optimal conditions for the AJ strain of HHV-6 were found to be culturing in HSB-2 cells at a concentration of 1:25-1:50 infected to uninfected cells and harvesting after 7 days.
为了优化病毒抗原的产生，在两种细胞系HSB - 2和JJHAN中研究了人类疱疹病毒6型（HHV - 6）（AJ株）的生长特性。用不同的感染复数（moi）感染细胞，并通过细胞病变效应（CPE）、总细胞计数和活细胞计数、免疫荧光试验、免疫印迹和电子显微镜观察来监测病毒生长。尽管高moi感染时>或= 70%的JJHAN细胞出现CPE，但免疫荧光检测时只有5%的细胞含有病毒抗原。相比之下，HSB - 2细胞在感染复数>或= 1:50时，出现荧光的细胞百分比达到>或= 30%。免疫印迹显示，HSB - 2培养物中出现了10多种分子量在31 - 140 kD之间的多肽，而在高moi的JJHAN培养物中仅检测到3种多肽。电子显微镜观察到HSB - 2细胞中病毒成熟的不同阶段，但病毒在JJHAN细胞中的复制似乎受到限制。为了生产抗原，发现HHV - 6 AJ株的最佳条件是在HSB - 2细胞中以1:25 - 1:50的感染复数感染未感染细胞，并在7天后收获。
