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一种利用细胞gp 71结合来监测小鼠白血病病毒感染的快速生物测定法。

A rapid bioassay to monitor murine leukemia virus infection in mice using cellular gp 71 binding.

作者信息

Reed C D, Fowler A K

出版信息

J Virol Methods. 1982 May;4(4-5):209-17. doi: 10.1016/0166-0934(82)90067-2.

Abstract

A rapid and sensitive bioassay based on the availability of cell surface receptors for the binding of purified envelope glycoprotein, gp71, of Rauscher murine leukemia virus (R-MuLV) was developed to serially monitor viral-induced leukemogenesis in individual BALB/cAnN mice. The specificity of the bioassay was demonstrated by the competition of [125I]gp71 cellular binding with murine ecotropic viruses, purified unlabelled R-MuLV envelope glycoprotein and by antiserum to R-MuLV gp71. In contrast, there was no effect on the [125I]gp71 binding level with the addition of murine xenotropic viruses, R-MuLV p30, or several other proteins. The [125I]gp71 binding level of circulating leukocytes was significantly (P less than 0.05) reduced in mice after R-MuLV infection. The reduction of cellular gp71 binding developed in two stages and the latter stage was highly dependent (P less than 0.05) on circulating infectious virus titer. Using this technique, the gp71 cellular binding levels of 48-60 individual mice can be assayed in a 4 h period. The advantages of this bioassay compared to standard immunological and tissue culture techniques used in studying retrovirus expression and viral-cell interactions are discussed.

摘要

基于劳斯氏鼠白血病病毒(R-MuLV)纯化包膜糖蛋白gp71与细胞表面受体结合的可用性,开发了一种快速灵敏的生物测定法,用于连续监测个体BALB/cAnN小鼠中病毒诱导的白血病发生。通过[125I]gp71细胞结合与鼠嗜亲性病毒、纯化的未标记R-MuLV包膜糖蛋白的竞争以及抗R-MuLV gp71血清,证明了该生物测定法的特异性。相比之下,添加鼠异嗜性病毒、R-MuLV p30或其他几种蛋白质对[125I]gp71结合水平没有影响。R-MuLV感染后,小鼠循环白细胞的[125I]gp71结合水平显著降低(P<0.05)。细胞gp71结合的降低分两个阶段发展,后一阶段高度依赖于循环感染性病毒滴度(P<0.05)。使用该技术,可在4小时内检测48至60只个体小鼠的gp71细胞结合水平。讨论了该生物测定法与用于研究逆转录病毒表达和病毒-细胞相互作用的标准免疫学和组织培养技术相比的优势。

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