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Structural analysis of envelope proteins of macrophage tropic HIV

嗜巨噬细胞HIV包膜蛋白的结构分析

基本信息

批准号：
14370159
负责人：
HATTORI Toshio
金额：
$ 7.3万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370159/
关键词：
gp41 HIV antibody AIDS vaccine gp120 ペプチドモノクロナル抗体立体構造 R5 X4 V3グループ fusogenic form raft エピトープ

项目摘要

To understand the mechanisms of HIV infection, we clarified that N terminus of V3 loop of BH-10 clone is responsible for its binding to the cells. We also treated the surface of H9/IIIB cells by several proteases including thrombin and analyzed the expression of functional epitomes by FACS. Thrombin treatment decreased the binding of anti-V3 mAb while enhanced the bindings of mAbs against Co-receptor binding site, a CD4 binding site and neutralizing epitope of gp41. Thrombin also enhanced fusion between H9/IIIB and MAGI cells. These findings support that blood borne inflammatory molecule thrombin activate gp 120 and enhanced the fusion.We also evaluated monoclonal antibodies (mAbs) against the gp41 core structure such as mAbs 50.69, 98.6 and T26, by western blotting and flow cytometry. Western blotting showed mAbs 50.69 and 98.6 bound to both monomeric and oligomeric gp41 and mAb T26 exclusively bound to oligomeric gp41. We evaluated the sera from pneumocystis pneumonia patients (PCP) and long term survivors (LTS). The results revealed that PCP sera as well as LTS sera inhibited the binding of all the three mAbs, and the PCP sera inhibited mAb T26 binding efficiently than LTS. Therefore PCP patients retain competing immunity to antibodies against not only the shared epitopes of the core structure (binding sites of mAbs 50.69 and 98.6) but also against oligomeric gp41 specific epitope (binding site of mAb T26).

为了了解HIV感染的机制，我们阐明了BH-10克隆的V3环的N末端负责其与细胞的结合。我们还用凝血酶等多种蛋白酶处理H9/IIIB细胞表面，并通过FACS分析功能表位的表达。凝血酶处理降低了抗 V3 mAb 的结合，同时增强了 mAb 对辅助受体结合位点、CD4 结合位点和 gp41 中和表位的结合。凝血酶还增强 H9/IIIB 和 MAGI 细胞之间的融合。这些发现支持血源性炎症分子凝血酶激活 gp 120 并增强融合。我们还通过蛋白质印迹和流式细胞术评估了针对 gp41 核心结构的单克隆抗体 (mAb)，例如 mAb 50.69、98.6 和 T26。 Western印迹显示mAb 50.69和98.6与单体和寡聚gp41结合，mAb T26仅与寡聚gp41结合。我们评估了肺孢子虫肺炎患者 (PCP) 和长期幸存者 (LTS) 的血清。结果显示，PCP 血清以及 LTS 血清均抑制所有三种 mAb 的结合，并且 PCP 血清比 LTS 更有效地抑制 mAb T26 结合。因此，PCP 患者保留了对抗体的竞争免疫，抗体不仅针对核心结构的共享表位（mAb 50.69 和 98.6 的结合位点），而且还针对寡聚 gp41 特异性表位（mAb T26 的结合位点）。