Preliminary studies on cell cultures infected with Marek's disease agent.
Preliminary studies on cell cultures infected with Marek's disease agent.
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对感染马立克氏病病原体的细胞培养物的初步研究。
DOI：
10.2307/1588098
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发表时间：
1968
期刊：
影响因子：
1.4
通讯作者：
J. Solomon
中科院分区：
文献类型：
--
作者：
R. Witter;G. Burgoyne;J. Solomon
目前，马立克氏病（MD）病原体的生物测定需要鸡接种，然后检查病变（2，10，13）。这些方法的有效性受到时间、易感鸟类和严格隔离设施的物理要求以及响应测量的主观性的限制。本文报道的研究是基于这样的前提，即细胞培养物中MD试剂的测定将构成对现有体内方法的显著改进。先前在细胞培养中繁殖MD试剂的尝试是不成功的或不确定的。Biggs和Payne（2）在第3代和第6代检测时，在接种B14菌株的鸡胚成纤维细胞培养物中未能证明感染性。Vindel（11）在细胞培养中繁殖，这是一种从患有神经淋巴瘤病的鸡中分离的滤过性因子。该试剂在鸡胚成纤维细胞中产生快速的细胞病变效应，并且用来自感染培养物的液体接种的30C鸡在3个月内发展为MD。然而，由于未描述接种雏鸡中发生的病变，并且接种物诱导病变的证据不确定，因此未明确确定该病原体的身份。尽管缺乏MD试剂在细胞培养中繁殖的证据，但细胞培养生物测定的潜在有用性似乎证明了进一步研究的合理性。本研究的最初目的是开发培养MD感染细胞的技术和用MD因子感染培养物的技术。本报告描述了成功地维持JM株的MD在鸡胚骨髓和某些其他细胞的培养。我们知道Calnek（4）和Kottarnik和Luginbuhl（7）同时进行的独立调查，但尚未公布，
Bioassay of Marek's disease (MD) agents presently requires chick inoculation and later examination for lesions (2,10,13). The usefulness of such methods is limited by the physical requirements of time, susceptible birds, and strict isolation facilities, and also by the subjectivity of the response measurement. The studies reported herein were based on the premise that the assay of MD agent in cell culture would constitute a significant improvement over existing in vivo methods. Previous attempts to propagate the MD agent in cell culture were either unsuccessful or equivocal. Biggs and Payne (2) failed to demonstrate infectivity in chick embryo fibroblast cultures inoculated with the B14 strain when tested at the 3rd and 6th passages. Vindel (11) propagated in cell culture a filtrable agent isolated from chickens with neurolymphomatosis. This agent produced a rapid cytopathic effect in chick embryo fibroblasts, and 30C of chickens inoculated with fluids from infected cultures developed MD within 3 months. The identity of this agent was not definitely established, however, since lesions occurring in the inoculated chicks were not described, and evidence that the lesions were induced by the inoculum was inconclusive. Despite the paucity of evidence for the propagation of MD agent in cell culture, the potential usefulness of a cell culture bioassay appeared to justify further investigation. The initial objective of this study was to develop techniques for cultivating MDinfected cells and for infecting cultures with MD agent. This report describes successful maintenance of the JM strain of MD in cultures of chick embryo bone marrow and certain other cells. We are aware of simultaneous and independent investigations by Calnek (4) and Kottaridis and Luginbuhl (7), as yet unpublished,