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支持1型和2型马立克氏病病毒以及火鸡疱疹病毒复制的鹌鹑细胞系。

Quail cell lines supporting replication of Marek's disease virus serotype 1 and 2 and herpesvirus of turkeys.

作者信息

Li Xinhui, Schat Karel Antoni

机构信息

Unit of Avian Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.

出版信息

Avian Dis. 2004 Dec;48(4):803-12. doi: 10.1637/7182-032604R.

Abstract

Marek's disease virus (MDV), a highly cell-associated alphaherpesvirus, can be isolated and propagated in chicken kidney cells (CKC) and chicken or duck embryo fibroblast cells (CEF or DEF, respectively). Two recently developed cell lines, CU447 and CU453, developed from methylcholanthrene-induced tumors in Japanese quail, were examined for their suitability to propagate the three serotypes of MDV. The MDV strain RB-1B (serotype 1) was passaged for more than 30 passages in CU447 without causing cytopathic effects (CPE). Polymerase chain reaction analysis of RB-1B-infected CU447 cells demonstrated the presence of MDV DNA using primers specific for ICP4, pp38, and gB. The 132-bp direct repeats within the BamH1-H and -D fragments were amplified to the same level as RB-1B that was passaged in CKC or CEF. Different passages of RB-1B in CU447 were examined for expression of gB and pp38 transcripts, and pp38, gB, gE, and VP22 protein expression. Irrespective of the passage level, these transcripts and proteins were detected in the RB-1B-infected CU447 cells. Infectious virus was rescued by cocultivation of RB-1B-infected CU447 with CKC. Herpes virus of turkeys was propagated in CU447 and CU453 causing CPE in both cell lines. SB-1 did not cause CPE in either cell line but a few SB-1-infected cells could be detected using a monoclonal antibody specific for serotype 2 MDV.

摘要

马立克氏病病毒(MDV)是一种高度细胞相关的α疱疹病毒,可在鸡肾细胞(CKC)以及鸡或鸭胚成纤维细胞(分别为CEF或DEF)中分离和增殖。对最近从甲基胆蒽诱导的日本鹌鹑肿瘤中培育出的两种细胞系CU447和CU453,检测了它们增殖MDV三种血清型的适用性。MDV毒株RB - 1B(血清1型)在CU447中传代30多次未引起细胞病变效应(CPE)。用针对ICP4、pp38和gB的特异性引物对感染RB - 1B的CU447细胞进行聚合酶链反应分析,证实存在MDV DNA。BamH1 - H和 - D片段内的132 bp直接重复序列扩增至与在CKC或CEF中传代的RB - 1B相同水平。检测了RB - 1B在CU447中的不同传代情况,以分析gB和pp38转录本的表达以及pp38、gB、gE和VP22蛋白的表达。无论传代水平如何,在感染RB - 1B的CU447细胞中均检测到这些转录本和蛋白。通过将感染RB - 1B的CU447与CKC共培养拯救出感染性病毒。火鸡疱疹病毒在CU447和CU453中增殖,在两种细胞系中均引起CPE。SB - 1在两种细胞系中均未引起CPE,但使用针对血清2型MDV的单克隆抗体可检测到少数感染SB - 1的细胞。

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