sCD4-17b bifunctional protein: extremely broad and potent neutralization of HIV-1 Env pseudotyped viruses from genetically diverse primary isolates.
sCD4-17b bifunctional protein: extremely broad and potent neutralization of HIV-1 Env pseudotyped viruses from genetically diverse primary isolates.
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sCD4-17b 双功能蛋白：对来自遗传多样性初级分离株的 HIV-1 Env 假型病毒具有极其广泛和有效的中和作用。
DOI：
10.1186/1742-4690-7-11
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发表时间：
2010-02-16
期刊：
影响因子：
3.3
通讯作者：
Berger EA
中科院分区：
文献类型：
--
作者：
Lagenaur LA;Villarroel VA;Bundoc V;Dey B;Berger EA
我们之前描述了一种有效的重组HIV-1中和蛋白，sCD4-17b，由可溶性CD4组成，通过柔性多肽接头连接到17b人单克隆抗体的SCFv上，该抗体针对高度保守的CD4诱导的gp120桥接片，参与共同受体结合。双功能蛋白的 sCD4 部分与游离病毒体上的 gp120 结合，从而使 17b SCFv 部分能够结合并阻断进入所需的 gp120/辅助受体相互作用。先前使用 MAGI-CCR5 检测系统的研究表明 sCD4-17b（在浓缩细胞培养基中，或部分纯化）可有效中和几种遗传多样性的 HIIV-1 原代分离株；然而，在测试的浓度下，尽管 CD4 和 17b 的结合位点得到保留，但它对其他几种菌株无效。为了解决这个难题，我们设计了具有不同接头长度的 sCD4-17b 变体，并使用 TZM-bl Env 假型检测系统在更广泛的浓度范围内测试了免疫亲和纯化蛋白对大量遗传多样性 HIV-1 原代分离株的中和活性。我们还检查了不同生产细胞类型产生的同基因病毒的 sCD4-17b 敏感性。我们观察到，免疫亲和纯化的 sCD4-17b 有效中和了 HIV-1 假型，包括先前在 MAGI-CCR5 测定中发现相对不敏感的 HIV-1 分离株。正如用假型颗粒和感染性病毒体观察到的那样，原始构建体和具有较长连接体的变体的效力相同；相比之下，连接子太短而无法同时结合 sCD4 和 17b SCFv 部分的构建体的效果要差得多。 sCD4-17b 对来自不同遗传亚型（进化枝 A、B、C、D、F 和循环重组形式 AE 和 AG）的近 4 打 HIV-1 原代分离株中的 100% 显示出有效的中和活性。中和广度和效力优于已报道的广泛中和单克隆抗体 IgG b12、2G12、2F5 和 4E10。发现 sCD4-17b 的活性与来自直接源自转染的生产细胞系或单次传代 PBMC 的感染性分子克隆的同基因病毒颗粒的活性相似；这与单克隆抗体形成鲜明对比，单克隆抗体对抗 PMBC 传代病毒的效力较差。结果强调了 sCD4-17b 对遗传多样性的 HIV-1 原代分离株具有极其有效和广泛的中和活性。这种双功能蛋白在对抗艾滋病毒感染的抗病毒方法中具有潜在的应用。
We previously described a potent recombinant HIV-1 neutralizing protein, sCD4-17b, composed of soluble CD4 attached via a flexible polypeptide linker to an SCFv of the 17b human monoclonal antibody directed against the highly conserved CD4-induced bridging sheet of gp120 involved in coreceptor binding. The sCD4 moiety of the bifunctional protein binds to gp120 on free virions, thereby enabling the 17b SCFv moiety to bind and block the gp120/coreceptor interaction required for entry. The previous studies using the MAGI-CCR5 assay system indicated that sCD4-17b (in concentrated cell culture medium, or partially purified) potently neutralized several genetically diverse HIIV-1 primary isolates; however, at the concentrations tested it was ineffective against several other strains despite the conservation of binding sites for both CD4 and 17b. To address this puzzle, we designed variants of sCD4-17b with different linker lengths, and tested the neutralizing activities of the immunoaffinity purified proteins over a broader concentration range against a large number of genetically diverse HIV-1 primary isolates, using the TZM-bl Env pseudotype assay system. We also examined the sCD4-17b sensitivities of isogenic viruses generated from different producer cell types. We observed that immunoaffinity purified sCD4-17b effectively neutralized HIV-1 pseudotypes, including those from HIV-1 isolates previously found to be relatively insensitive in the MAGI-CCR5 assay. The potencies were equivalent for the original construct and a variant with a longer linker, as observed with both pseudotype particles and infectious virions; by contrast, a construct with a linker too short to enable simultaneous binding of the sCD4 and 17b SCFv moieties was much less effective. sCD4-17b displayed potent neutralizing activity against 100% of nearly 4 dozen HIV-1 primary isolates from diverse genetic subtypes (clades A, B, C, D, F, and circulating recombinant forms AE and AG). The neutralization breadth and potency were superior to what have been reported for the broadly neutralizing monoclonal antibodies IgG b12, 2G12, 2F5, and 4E10. The activity of sCD4-17b was found to be similar against isogenic virus particles from infectious molecular clones derived either directly from the transfected producer cell line or after a single passage through PBMCs; this contrasted with the monoclonal antibodies, which were less potent against the PMBC-passaged viruses. The results highlight the extremely potent and broad neutralizing activity of sCD4-17b against genetically diverse HIV-1 primary isolates. The bifunctional protein has potential applications for antiviral approaches to combat HIV infection.
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影响因子：
4.9
作者：
Liu, Xiaowen;Lagenaur, Laurel A.;Xu, Qiang
通讯作者：
Xu, Qiang
影响因子：
5.4
作者：
Dey, B;Del Castillo, CS;Berger, EA
通讯作者：
Berger, EA
影响因子：
64.5
作者：
Choe, H;Li, WH;Farzan, M
通讯作者：
Farzan, M
DOI：
10.1073/pnas.0707399104
发表时间：
2007-10-30
影响因子：
11.1
作者：
DeVico, Anthony;Fouts, Timothy;Pal, Ranajit
通讯作者：
Pal, Ranajit
国际艾滋病毒中和试验比较网络：NeutNet 报告。
影响因子：
3.7
作者：
Fenyo, Eva Maria;Heath, Alan;Dispinseri, Stefania;Holmes, Harvey;Lusso, Paolo;Zolla-Pazner, Susan;Donners, Helen;Heyndrickx, Leo;Alcami, Jose;Bongertz, Vera;Jassoy, Christian;Malnati, Mauro;Montefiori, David;Moog, Christiane;Morris, Lynn;Osmanov, Saladin;Polonis, Victoria;Sattentau, Quentin;Schuitemaker, Hanneke;Sutthent, Ruengpung;Wrin, Terri;Scarlatti, Gabriella
通讯作者：
Scarlatti, Gabriella