Animal, Plant and Fisheries Quarantine and Inspection Agency, Jungbu Regional Office, Incheon 400-800, Republic of Korea.
J Microbiol. 2012 Oct;50(5):821-6. doi: 10.1007/s12275-012-2364-2. Epub 2012 Nov 4.
Infectious pancreatic necrosis virus (IPNV), a type species of aquabirnaviruses in the family Birnaviridae, is an etiological agent of infectious pancreatic necrosis and has been isolated from epizootics of cultured salmonids. In the present study, an epithelioma papulosum cyprini (EPC) cell line persistently infected with IPNV (PI-EPC) was experimentally established by subculturing EPC cells surviving IPNV infection, and was characterized. PI-EPC cells were morphologically indistinguishable from EPC, but continued to grow and yield IPNV. PI-EPC cells showed no cytopathic effect due to IPNV inoculation, and susceptibility of PI-EPC cells against heterologous viruses was not different from that of EPC cells. Only one cell of 10(3.5) PI-EPC cells produced IPNV at approximately 10(0.5) 50% tissue culture infectious dose (TCID50)/cell/day, which was approximately 1,000 times lower than that of normal EPC cells. PI-EPC cells that did not yield IPNV (N-PI-EPC) were screened. The IPNV genome was detected from both PI-EPC and N-PI-EPC cells, and the IPNV VP2 structural protein was detected from both cell lines, but no other IPNV proteins were observed by Western blot analysis with anti-IPNV serum. Thus, multiplication of IPNV in PI-EPC cells was regulated by some host cell factors, except interferon.
传染性胰脏坏死病毒(IPNV),属于双RNA 病毒科双RNA 病毒属的一个种,是传染性胰脏坏死的病原体,已从养殖鲑鱼的疫病中分离得到。在本研究中,通过传代感染 IPNV 后存活的上皮瘤细胞(EPC),实验建立了一株持续感染 IPNV 的 EPC 细胞(PI-EPC)系,并对其进行了特征分析。PI-EPC 细胞在形态上与 EPC 细胞无明显区别,但能持续生长并产生 IPNV。由于接种了 IPNV,PI-EPC 细胞没有出现细胞病变效应,且对异源病毒的敏感性与 EPC 细胞无差异。在大约 10(0.5) 50%组织培养感染剂量(TCID50)/细胞/天时,只有 10(3.5)个 PI-EPC 细胞中的一个细胞能产生 IPNV,这大约是正常 EPC 细胞的 1000 倍。筛选出未产生 IPNV 的 PI-EPC 细胞(N-PI-EPC)。PI-EPC 和 N-PI-EPC 细胞均检测到了 IPNV 基因组,且这两种细胞系均检测到了 IPNV VP2 结构蛋白,但用抗 IPNV 血清进行 Western blot 分析时未观察到其他 IPNV 蛋白。因此,除干扰素外,宿主细胞的某些因子调节了 IPNV 在 PI-EPC 细胞中的增殖。