Amadori M, Volpe G, Defilippi P, Berneri C
Istituto Zooprofilattico Sperimentale, Department of Vaccine Research and Development, Brescia, Italy.
Biologicals. 1997 Mar;25(1):65-73. doi: 10.1006/biol.1996.0061.
BHK-21 c13 monolayer and suspension cells were investigated with regard to some phenotypic features which could bear on the quality of foot-and-mouth disease virus (FMDV) antigen produced in them. Despite good viability, suspension cells differed from monolayer cells in fundamental features of susceptibility to FMDV. Most important, FMD virus particles grown in suspension cells at high passage levels were shown to be largely degraded following inactivation with an aziridine compound. Suspension cells were characterised by a downregulation in the surface expression of both alpha 5 and alpha V integrin chains. According to the results of binding assays, both integrins could act as FMDV receptors on BHK-21 c13 cells. Reduced surface expression of integrins was correlated with disappearance of actin stress fibres, which would play a role in regular encapsidation of viral RNA and hence in stability of virus particles. With regard to FMD vaccine production, practical suggestions are put forward to evaluate the quality of BHK-21 c13 cells and FMDV Ag, which must prove stable during downstream processing.
对BHK - 21 c13单层细胞和悬浮细胞的一些表型特征进行了研究,这些特征可能与它们产生的口蹄疫病毒(FMDV)抗原的质量有关。尽管悬浮细胞活力良好,但在对FMDV的易感性基本特征方面与单层细胞不同。最重要的是,高传代水平下在悬浮细胞中生长的FMD病毒颗粒在用氮丙啶化合物灭活后显示出大量降解。悬浮细胞的特征是α5和αV整合素链的表面表达下调。根据结合试验结果,两种整合素都可作为BHK - 21 c13细胞上的FMDV受体。整合素表面表达的降低与肌动蛋白应力纤维的消失相关,肌动蛋白应力纤维在病毒RNA的正常包装中起作用,从而在病毒颗粒的稳定性中起作用。关于FMD疫苗生产,提出了实际建议,以评估BHK - 21 c13细胞和FMDV Ag的质量,它们在下游加工过程中必须保持稳定。