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Assays for Endogenous and Exogenous Lymphoid Leukosis Viruses and Chick Helper Factor with RSV(−) Cell Lines

机译:RSV(-)细胞系的内源性和外源性淋巴白血病病毒和雏鸡辅助因子测定

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摘要

Japanese quail cells transformed by the envelope-defective Bryan high-titer strain of Rous sarcoma virus [R(−)Q] were used as a source of the Rous sarcoma virus genome in three kinds of assays. (i) The simplest and most sensitive assay for infectious, endogenous viruses of the chicken belonging to subgroup E involved infection of a mixture of R(−)Q cells and turkey cells with the sample and assay of supernatants of these cells for focus formation on subgroup E susceptible cells. (ii) Inactivated Sendai virus-induced fusion of R(−)Q cells with live test cells was found to be a specific method for detection of chick helper factor. Focus formation by supernatant of the fused cells on subgroup E susceptible cells was correlated with the presence of subgroup E envelope glycoprotein on the plasma membranes of test cells. Whole blood cells as well as fibroblasts could be used in this assay. (iii) A method of assay for exogenous lymphoid leukosis viruses in which mixed cultures of R(−)Q cells and C/E cells and assay of supernatants for focus formation on C/E cells was as sensitive as assays presently used for exogenous lymphoid leukosis virus. Because no infectious Rous sarcoma virus was used as part of the procedure, the assays for infectious virus described here yielded pure pseudotypes of the input virus, an advantage for determining purity and subgroup of the input virus.
机译:在三种检测方法中,将由包膜缺陷性布劳恩高滴度劳斯肉瘤病毒[R(-)Q]转化的日本鹌鹑细胞用作劳斯肉瘤病毒基因组的来源。 (i)最简单,最灵敏的方法是对属于E亚组的鸡的传染性,内源性病毒进行检测,包括用样品感染R(-)Q细胞和火鸡细胞的混合物,并检测这些细胞的上清液以形成斑点。 E亚群易感细胞。 (ii)灭活的仙台病毒诱导的R(-)Q细胞与活细胞的融合是发现雏鸡辅助因子的一种特定方法。融合细胞上清液在E组亚型易感细胞上形成的病灶形成与测试细胞质膜上E组亚膜糖蛋白的存在有关。全血细胞以及成纤维细胞可以用于该测定中。 (iii)一种外源性淋巴白血病病毒的测定方法,其中R(-)Q细胞和C / E细胞的混合培养物以及用于在C / E细胞上形成焦点的上清液的测定方法与目前用于外源性淋巴瘤的测定方法一样灵敏白血病病毒。因为没有传染性劳斯肉瘤病毒被用作该程序的一部分,所以此处描述的传染性病毒测定法可产生纯的输入病毒假型,这对于确定输入病毒的纯度和亚组是一个优势。

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