N-butyldeoxynojirimycin-mediated inhibition of human immunodeficiency virus entry correlates with changes in antibody recognition of the V1/V2 region of gp120
N-butyldeoxynojirimycin-mediated inhibition of human immunodeficiency virus entry correlates with changes in antibody recognition of the V1/V2 region of gp120
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DOI：
10.1128/jvi.70.10.7143-7152.1996
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发表时间：
1996-10-01
影响因子：
5.4
通讯作者：
Platt, FM
中科院分区：
文献类型：
--
作者：
Fischer, PB;Karlsson, GB;Platt, FM
α-葡萄糖苷酶抑制剂 N-丁基脱氧野尻霉素 (NB-DNJ) 是人类免疫缺陷病毒 (HIV) 复制和 HIV 诱导的体外合胞体形成的抑制剂，尽管 NB-T)NJ 似乎在 CD4 结合后水平上抑制 HIV 进入（P. B. Fischer、M. Collin、G. B. Karlsson、W. James、T. D. Butters、S. J. Davis、S. Gordon、R. A. Dwek 和 F. M. Platt、J. Virol。 69:5791-5797, 1995），确切的作用机制仍有待确定，在这项研究中，我们通过使用面板检查了 KB-DNJ 对 CHO 细胞中表达的重组 gp120 (rgp120) 结构的影响 40种单克隆抗体，通过酶联免疫吸附测定和表面等离子共振（BIAcore；Pharmacia）比较了存在[rgp120（+）]和不存在[rgp120（-）]NB-DNJ时产生的抗体与rgp120的结合水平，结果显示与rgp120（+）的结合增加 与结合 rgp120(-) 的抗体相比，针对 C1 和 C2 区域的抗体和针对 V1/V2 环的抗体的结合减少，还观察到针对不连续表位的抗体与 rgp120(+) 的结合减少，针对 V3 环冠的抗体的结合没有看到差异 和 gp120 的 C4 区域，用 α-葡萄糖苷酶 I 和 II 处理 rgp120 对观察到的差异结合没有影响，而用唾液酸酶处理消除了针对 gp120 C1 和 C2 区域的抗体结合中观察到的差异。除了这些发现之外，rgp120(+) 对 CHO 细胞在表达过程中释放的蛋白酶的敏感性也增加 对于外源性凝血酶，综上所述，本文提供的数据表明，在 NB-DNJ 存在的情况下 gp120 的产生会影响 gp120 V1/V2 环的构象，以及 C1 和 C2 区域的总电荷。这些效应可能在先前描述的 NB-DNJ 介导的 CD4 结合后水平的 HIV 进入抑制中发挥作用。
The alpha-glucosidase inhibitor N-butyldeoxynojirimycin (NB-DNJ) is an inhibitor of human immunodeficiency virus (HIV) replication and HIV-induced syncytium formation in vitro, Although NB-T)NJ appears to inhibit HIV entry at the level of post-CD4 binding (P. B. Fischer, M. Collin, G. B. Karlsson, W. James, T. D. Butters, S. J. Davis, S. Gordon, R. A. Dwek, and F. M. Platt, J. Virol. 69:5791-5797, 1995), the exact mechanism of action remains to be established, In this study we have examined the effect of KB-DNJ on the structure of recombinant gp120 (rgp120), expressed in CHO cells, by using a panel of 40 monoclonal antibodies, The levels of binding of antibodies to rgp120 produced in the presence [rgp120(+)] and absence [rgp120(-)] of NB-DNJ were compared by enzyme-linked immunosorbent assay and surface plasmon resonance (BIAcore; Pharmacia), The results showed an increase in the binding to rgp120(+) of antibodies directed against the C1 and C2 regions and a decrease in the binding of antibodies directed against the V1/V2 loops compared with antibody binding to rgp120(-), A decrease in the binding to rgp120(+) of antibodies directed against discontinuous epitopes was also observed, No differences were seen in the binding of antibodies directed against the crown of the V3 loop and the C4 region of gp120, Treatment of rgp120 with alpha-glucosidases I and II had no effect on the differential binding observed, whereas treatment with sialidase abolished the differences seen in the binding of antibodies directed against the C1 and C2 regions of gp120, In addition to these findings, rgp120(+) showed increased sensitivity to proteases released by CHO cells during expression, as well as to exogenous thrombin, Taken together, the data presented in this paper suggest that production of gp120 in the presence of NB-DNJ affects the conformation of the V1/V2 loops of gp120, as well as the overall charge of the C1 and C2 regions. These effects may play a role in the previously described NB-DNJ-mediated inhibition of HIV entry at the level of post-CD4 binding.