Rong Sing, Wheeler David, Weber Fred
a Global Biologics Research, Veterinary Medicine Research & Development, Zoetis, Kalamazoo , Michigan , USA.
Avian Pathol. 2014;43(5):414-9. doi: 10.1080/03079457.2014.956687.
Marek's disease virus (MDV; also known as Gallid herpesvirus 2, MDV-1) causes oncogenic disease in chickens producing clinical signs that include lymphomas, visceral tumours, nerve lesions, and immunosuppression. MDV vaccines are widely used and mostly produced using primary cells: chicken embryo fibroblast cells, duck embryo fibroblast cells, chicken embryo kidney cells or chicken kidney cells. An immortalized cell line that can be used to manufacture the virus has long been desired. In this report, we demonstrate that QM7 cells were susceptible to infection with either MDV or herpesvirus of turkey (HVT; also known as Meleagrid herpesvirus 1, MDV-3). Polymerase chain reaction analysis with primers amplifying selected MDV genes revealed that QM7 cells did not contain these sequences. However, MDV genes were detected in QT35 cells, which have been reported to harbour latent MDV virus. Transfection of naked MDV DNA initiated efficient infection of QM7 cells. In addition, QM7 cell lysate, clarified supernatant, and QM7 cell pellet infected with MDV were negative for reverse transcriptase activity, indicating absence of endogenous retrovirus. QM7 cells were also found to be free of other avian pathogens in a chick embryo inoculation test. In vivo studies of MDV growing in QM7 cells showed the virus retained its pathogenicity and virulence. In ovo experiments demonstrated that both HVT and MDV propagated in QM7 cells did not interfere with hatchability of injected eggs, and viruses could be re-isolated from hatched chicks. The results suggest that QM7 could be a good host cell line for growing both MDV and HVT.
马立克氏病病毒(MDV;也称为鸡疱疹病毒2型,MDV - 1)可导致鸡发生致癌性疾病,产生包括淋巴瘤、内脏肿瘤、神经病变和免疫抑制等临床症状。MDV疫苗被广泛使用,且大多使用原代细胞生产:鸡胚成纤维细胞、鸭胚成纤维细胞、鸡胚肾细胞或鸡肾细胞。长期以来一直期望有一种可用于生产该病毒的永生化细胞系。在本报告中,我们证明QM7细胞易受MDV或火鸡疱疹病毒(HVT;也称为火鸡疱疹病毒1型,MDV - 3)感染。用扩增选定MDV基因的引物进行聚合酶链反应分析表明,QM7细胞不包含这些序列。然而,在已报道含有潜伏MDV病毒的QT35细胞中检测到了MDV基因。裸MDV DNA转染引发了QM7细胞的有效感染。此外,感染MDV的QM7细胞裂解物、澄清上清液和QM7细胞沉淀的逆转录酶活性均为阴性,表明不存在内源性逆转录病毒。在鸡胚接种试验中还发现QM7细胞不含其他禽病原体。对在QM7细胞中生长的MDV进行的体内研究表明,该病毒保留了其致病性和毒力。卵内实验表明,在QM7细胞中增殖的HVT和MDV均不影响注射卵的孵化率,并且可以从孵化出的雏鸡中重新分离出病毒。结果表明,QM7可能是用于培养MDV和HVT的良好宿主细胞系。