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Human T-cell line infected with HIV-2 which secretes a protein corresponding to native HIV-2 gp160 in an extracellular medium

机译:感染HIV-2的人T细胞系在细胞外培养基中分泌与天然HIV-2 gp160相对应的蛋白质

摘要

A novel cell line chronically infected with HIV-2.sub.NIHZ virus secretes the precursor of the viral envelope proteins in extracellular medium. The cells grow well in serum-free medium which enables the concentration of the conditioned medium of the cells to be greatly increased ( 100 fold). Such a concentration step is otherwise impossible to achieve if the cells are grown under conventional conditions using 10% fetal calf serum. A specific affinity procedure is provided to purify the gp160 protein from the concentrated conditioned medium of cultures of 100 liter batches using a mouse monoclonal antibody to the HIV-2 gp41 protein, which is a part of the gp160 protein. This purified HIV-2 gp160 reacts with high specificity and sensitivity with the anti viral antibodies of HIV-2 infected humans by several criteria such as i) ELISA, ii) western blot analysis, and iii) radioimmunoprecipitation of metabolically labeled gp160 from the same cell line. The purified HIV-2 gp160 from this cell line is therefore a diagnostic tool for the detection of HIV-2 infection in humans. In view of the predicted oligomeric property of gp160 and its close conformational similarity to the virion envelope, the purified gp160 can also be useful for the development of vaccine strategies against HIV-2 in humans.
机译:长期感染HIV-2.NIHZ病毒的​​新型细胞系在细胞外培养基中分泌病毒包膜蛋白的前体。细胞在无血清培养基中生长良好,这可使细胞条件培养基的浓度大大增加(> 100倍)。如果细胞在常规条件下使用10%胎牛血清生长,则不可能实现这种浓缩步骤。提供了一种特异性亲和程序,使用针对HIV-2 gp41蛋白的小鼠单克隆抗体从100升批次的浓缩培养液中纯化gp160蛋白,该抗体是gp160蛋白的一部分。这种纯化的HIV-2 gp160与HIV-2感染的人的抗病毒抗体具有高特异性和敏感性,可通过多种标准进行反应,例如i)ELISA,ii)免疫印迹分析和iii)放射性免疫沉淀来自同一细胞的代谢标记gp160线。因此,从该细胞系纯化的HIV-2 gp160是检测人类HIV-2感染的诊断工具。鉴于gp160的预测寡聚特性及其与病毒体被膜的紧密构象相似性,纯化的gp160也可用于开发针对人类的HIV-2疫苗策略。

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