Zhu Yu, Lau Adeline, Lau Jennifer, Jia Qiang, Karuppannan Anbu K, Kwang Jimmy
Animal Health Biotechnology Group, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, 117604, Singapore.
Virology. 2007 Dec 20;369(2):423-30. doi: 10.1016/j.virol.2007.08.014. Epub 2007 Sep 24.
Post-weaning multi-systemic wasting syndrome (PMWS) has emerged as a major disease that poses a significant threat to the economics of global swine industry. Porcine circovirus type 2 (PCV2) is the causal agent of PMWS in pigs. Currently, the prevention of PCV2 infection based on vaccines is limited, and the available vaccines are either killed viral vaccines or recombinant protein based vaccines and not cost effective. The PK-15 cells, which is widely used for PCV2 propagation, is not efficient and heterogeneous in terms of permissivity to viral infection. In order to acquire a homogeneous porcine kidney cell line that can reliably produce PCV2 in high titers, cell clones that show high- (PK15-C1) or low-permissive (PK15-A2) phenotype to PCV2 infection were derived from heterogeneous PK15 parent cells by limiting dilution and cell cloning. Maximum virus titers in PK15-C1, PK15-A2 and PK15 parent cells were 10(8), 10(2) and 10(5) tissue culture infective dose 50 (TCID 50)/ml, respectively. The viral proteins of PCV2 were produced and accumulated faster in PK15-C1 cells than those in PK15 parent cells. These results indicate that PK15-C1 cell clone is more permissive to PCV2 infection than PK15 parent cells and thus will be useful for PCV2 replication in vitro, as well as, vaccines, diagnostic and research applications on PCV2.
断奶后多系统消耗综合征(PMWS)已成为对全球养猪业经济构成重大威胁的主要疾病。猪圆环病毒2型(PCV2)是猪PMWS的病原体。目前,基于疫苗预防PCV2感染的效果有限,现有的疫苗要么是灭活病毒疫苗,要么是基于重组蛋白的疫苗,且成本效益不高。广泛用于PCV2增殖的PK-15细胞在病毒感染的允许性方面效率不高且具有异质性。为了获得能够可靠地高滴度产生PCV2的均匀猪肾细胞系,通过有限稀释和细胞克隆从异质的PK15亲本细胞中获得了对PCV2感染表现出高允许性(PK15-C1)或低允许性(PK15-A2)表型的细胞克隆。PK15-C1、PK15-A2和PK15亲本细胞中的最大病毒滴度分别为10^8、10^2和10^5组织培养感染剂量50(TCID50)/ml。PCV2的病毒蛋白在PK15-C1细胞中比在PK15亲本细胞中产生和积累得更快。这些结果表明,PK15-C1细胞克隆比PK15亲本细胞对PCV2感染更敏感,因此将有助于PCV2的体外复制以及PCV2的疫苗、诊断和研究应用。