Morozova O V, Grishechkin A E, Bakhvalova V N, Isaeva E I, Podcherniaeva R Ia
Vopr Virusol. 2012 Mar-Apr;57(2):40-3.
The currently used tick-borne encephalitis virus vaccines are based on the inactivation of tick-borne encephalitis virus (TBEV) of Far Eastern or West European genetic types from the primary cultures of chick embryo fibroblasts. Since the WHO recommends that vaccines should be designed using continuous cell cultures rather than chick embryos as a substrate, this investigation has compared the infection of continuous monolayer SPEV, Vero E6, and vaccine line Vero (B) cell cultures with TBEV strains of the Siberian and Far Eastern genetic types dominating in the endemic regions of Russia. After cell infection with Far Eastern (Sofyin and 205 strains) or Siberian (Aina, 2530, 2689, and 2703 strains) TBEV genetic types, the viable TBEV titers reached 2.8 Ig CPD50 for Vero (B) cells, 5.5 Ig CPD50 for Vero E6 cells, and up to 9 Ig CPD50 for SPEV cells. The quantitative scores of TBEV E antigen in enzyme immunoassay (EIA) and genome equivalents by reverse-transcription polymerase chain reaction (PCR), followed by real-time PCR, permitted one to estimate as high as 108 virions in 1 ml of culture fluid, which corresponded to those of the microscopic observations of CPD for SPEV cells and substantially exceeded the values for Vero E6 cells, and for Vero (B) cells in particular. The data of TBEV strain titration, EIA, and realtime reverse-transcription PCR suggest that the Russian vaccine Vero (B) cell line defined as meeting the WHO requirements, as well as Vero E6 cells may be used to design tick-borne encephalitis vaccine.
目前使用的蜱传脑炎病毒疫苗是基于对来自鸡胚成纤维细胞原代培养物的远东或西欧基因类型的蜱传脑炎病毒(TBEV)进行灭活。由于世界卫生组织建议疫苗应使用连续细胞培养物而非鸡胚作为底物来设计,本研究比较了连续单层SPEV、Vero E6和疫苗株Vero(B)细胞培养物被在俄罗斯流行地区占主导地位的西伯利亚和远东基因类型的TBEV毒株感染的情况。在用远东(索芬和205株)或西伯利亚(艾娜、2530、2689和2703株)TBEV基因类型感染细胞后,Vero(B)细胞的活TBEV滴度达到2.8 Ig CPD50,Vero E6细胞为5.5 Ig CPD50,SPEV细胞高达9 Ig CPD50。通过酶免疫测定(EIA)对TBEV E抗原进行定量评分,并通过逆转录聚合酶链反应(PCR),随后进行实时PCR测定基因组当量,使得能够估计出每毫升培养液中高达108个病毒粒子,这与SPEV细胞CPD的显微镜观察结果相对应,并且大大超过了Vero E6细胞,尤其是Vero(B)细胞的值。TBEV毒株滴定、EIA和实时逆转录PCR的数据表明,被定义为符合世界卫生组织要求的俄罗斯疫苗Vero(B)细胞系以及Vero E6细胞可用于设计蜱传脑炎疫苗。