Yoshikawa Rokusuke, Sato Eiji, Miyazawa Takayuki
Laboratory of Signal Transduction, Department of Cell Biology, Institute for Virus Research, Kyoto University, 53 Shogoin-Kawaracho, Sakyo-ku, Kyoto 606-8507, Japan.
Biologicals. 2011 Jan;39(1):33-7. doi: 10.1016/j.biologicals.2010.11.001. Epub 2010 Dec 8.
All domestic cats have a replication-competent endogenous retrovirus, termed RD-114 virus, in their genome and several feline cell lines produce RD-114 viruses. Recently, we found that a portion of live attenuated feline and canine vaccines produced using feline cell lines was contaminated with infectious RD-114 viruses. In this study, we expanded our survey and examined canine vaccines produced using 'non-feline' cell lines. Consequently, we found two vaccines containing RD-114 viral RNA by reverse transcriptase (RT)-polymerase chain reaction (PCR) and real-time RT-PCR. We also confirmed the presence of infectious RD-114 virus in the vaccines by the LacZ marker rescue assay and PCR to detect proviral DNA in TE671 cells (human rhabdomyosarcoma cells) inoculated with the vaccines. It is impossible to investigate the definitive cause of contamination with RD-114 virus; however, we suspect that a seed canine parvovirus type 2 was contaminated with RD-114 virus, because many canine parvoviruses have been isolated and attenuated using feline cell lines. To exclude RD-114 virus from live attenuated vaccines, we must pay attention to the contamination of seed viruses with RD-114 virus in addition to avoiding feline cell lines producing RD-114 virus when manufacturing vaccines.
所有家猫的基因组中都有一种具有复制能力的内源性逆转录病毒,称为RD - 114病毒,并且几种猫科细胞系会产生RD - 114病毒。最近,我们发现部分使用猫科细胞系生产的减毒猫用和犬用活疫苗被传染性RD - 114病毒污染。在本研究中,我们扩大了调查范围,检测了使用“非猫科”细胞系生产的犬用疫苗。结果,我们通过逆转录酶(RT)-聚合酶链反应(PCR)和实时RT - PCR发现两种疫苗含有RD - 114病毒RNA。我们还通过LacZ标记拯救试验和PCR确认了疫苗中存在传染性RD - 114病毒,以检测接种疫苗的TE671细胞(人横纹肌肉瘤细胞)中的前病毒DNA。目前无法调查RD - 114病毒污染的确切原因;然而,我们怀疑2型犬细小病毒种子被RD - 114病毒污染,因为许多犬细小病毒是使用猫科细胞系分离和减毒的。为了从减毒活疫苗中排除RD - 114病毒,除了在生产疫苗时避免使用产生RD - 114病毒的猫科细胞系外,我们还必须注意种子病毒被RD - 114病毒污染的情况。