[HIV-1 gp60在V3环部分或完全缺失的克隆、表达和表征]。

C Lavallée, L Thibodeau
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引用次数: 0

摘要

为了更好地理解免疫显性V3环在类型特异性免疫应答中的作用,并确定该序列是否在艾滋病发病机制中起作用,特别是在诱导CD4+细胞凋亡中,我们引入了pNL4-3中env基因的2个修饰:在V3环部分缺失,只保留环GPGRAF一致序列的保守尖端(env delta V3-GPGRAF),其次,V3序列加上C3中的43个核苷酸的完全缺失(env delta V3+)。这些结构以及未修饰的env基因被克隆并在杆状病毒系统中表达。Western blot分析表明,这两种修饰的env基因产物与参考抗hiv -1血清的反应程度与未修饰的gp 160相同。然而,与未修饰的env蛋白和env delta V3- gpgraf相比,虽然V3不直接参与受体结合,但env delta V3+蛋白不能与CD4分子结合。这些修饰和未修饰的重组蛋白将非常有助于确定部分或全部V3缺失的gp 160诱导广泛反应性中和抗体的潜力,以及确定V3是否在hiv诱导的某些发病机制中起作用,特别是细胞凋亡。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Cloning, expression and characterization of HIV-1 gp60 partially or completely deleted in the V3 loop].

In order to better understand the role of the immunodominant V3 loop in the type-specific immune response and also to determine if this sequence has a role in AIDS pathogenesis, notably in the induction of apoptosis in CD4+ cells, we have introduced 2 modifications in the env gene from pNL4-3: a partial deletion in the V3 loop, keeping only the conserved tip of the loop GPGRAF consensus sequence (env delta V3-GPGRAF) and, secondly, a complete deletion of V3 sequence plus 43 nucleotides in C3 (env delta V3+). These constructions as well as the non-modified env gene, were cloned and expressed in a baculovirus system. Western blot analysis has shown that both modified env gene products reacted with a reference anti-HIV-1 serum to the same extent as the non-modified gp 160. However, in contrast to the non-modified env-protein and to env delta V3-GPGRAF, the env delta V3+ protein failed to bind to CD4 molecule, although V3 is not directly involved in receptor binding. These modified and non-modified recombinant proteins will be very useful to determine the potential of the partially or totally V3-deleted gp 160 to induce broadly reactive neutralizing antibodies and also to determine if V3 has a role in certain aspects of HIV-induced pathogenesis, notably apoptosis.

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