Flores E F, Donis R O
Department of Veterinary and Biomedical Sciences, IANR, University of Nebraska-Lincoln 68583-0905, USA.
Virology. 1995 Apr 20;208(2):565-75. doi: 10.1006/viro.1995.1187.
A cell line, termed CRIB, resistant to infection with bovine viral diarrhea virus (BVDV) has been derived from the MDBK bovine kidney cell line. CRIB cells were obtained by selection and cloning of cells surviving infection with a highly cytolytic BVDV strain. CRIB cells contain no detectable infectious or defective BVDV as ascertained by cocultivation, animal inoculation, indirect immunofluorescence, Western immunoblot, Northern hybridization, and RNA PCR. Inoculation of CRIB cells with 24 cytopathic and noncytopathic BVDV strains does not result in expression of viral genes or amplification of input virus. Karyotype and isoenzyme analyses demonstrated that CRIB are genuine bovine cells. CRIB cells are as susceptible as the parental MDBK cells to 10 other bovine viruses, indicating that these cells do not have a broad defect blocking viral replication. Transfection of CRIB cells with BVDV RNA or virus inoculation in the presence of polyethylene-glycol results in productive infection, indicating that the defect of CRIB cells is at the level of virus entry. CRIB cells are the first bovine cells reported to be resistant to BVDV infection in vitro and may be a useful tool for studying the early interactions of pestiviruses with host cells.
一种名为CRIB的细胞系已从MDBK牛肾细胞系中获得，该细胞系对牛病毒性腹泻病毒（BVDV）感染具有抗性。CRIB细胞是通过对感染高细胞溶解性BVDV毒株后存活的细胞进行筛选和克隆而获得的。通过共培养、动物接种、间接免疫荧光、Western免疫印迹、Northern杂交和RNA PCR确定，CRIB细胞中未检测到可感染的或有缺陷的BVDV。用24种细胞病变和非细胞病变的BVDV毒株接种CRIB细胞不会导致病毒基因表达或输入病毒的扩增。核型和同工酶分析表明CRIB是真正的牛细胞。CRIB细胞与亲本MDBK细胞一样易受其他10种牛病毒的感染，这表明这些细胞没有广泛的缺陷来阻止病毒复制。用BVDV RNA转染CRIB细胞或在聚乙二醇存在下接种病毒会导致有效感染，这表明CRIB细胞的缺陷在于病毒进入水平。CRIB细胞是首次报道的在体外对BVDV感染具有抗性的牛细胞，可能是研究瘟病毒与宿主细胞早期相互作用的有用工具。
