Well-ordered trimeric HIV-1 subtype B and C soluble spike mimetics generated by negative selection display native-like properties.
Well-ordered trimeric HIV-1 subtype B and C soluble spike mimetics generated by negative selection display native-like properties.
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DOI：
10.1371/journal.ppat.1004570
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发表时间：
2015-01
期刊：
影响因子：
6.7
通讯作者：
Wyatt RT
中科院分区：
文献类型：
--
作者：
Guenaga J;de Val N;Tran K;Feng Y;Satchwell K;Ward AB;Wyatt RT
BG 505 gp 140 SOSIP是天然HIV-1包膜糖蛋白（Env）的可溶性模拟物，其结构标志着基于Env结构的免疫原设计新时代的开始。这些A亚型衍生的三聚体显示出有序的四级结构，显示出出色的抗原谱，可区分广泛中和抗体（bNAb）和非广泛中和抗体（non-bNAb）的识别，并提供坚实的基于Env的免疫原性平台起点。即使有了这一重要进展，获得来自其他亚型的均匀有序的可溶性SOSIP三聚体仍然具有挑战性。在这里，我们报告了“拯救”同质有序亚型B和C SOSIP三聚体从异质性Env混合物中使用CD 4结合位点定向（CD 4 bs）的非bNAbs在负选择纯化过程。这些非bNAb仅识别无序三聚体上的初级受体CD 4 b，但由于空间位阻而不识别天然Env刺突或良好有序的可溶性三聚体上的初级受体CD 4 b。阴性选择去除无序的低聚物后，我们证明了良好的秩序，均匀的三聚体的电子显微镜（EM）的恢复。我们获得了未配体的三聚体以及与sCD 4复合的3D EM重建，一组CD 4 bs定向的bNAb和切割依赖性三聚体特异性bNAb，PGT 151。使用生物层光干涉法（BLI），我们证明了有序的三聚体被bNAb有效地识别，而被非bNAb识别得很差，代表了天然病毒刺突的可溶性模拟物。生物物理表征与可以通过特定bNAb进一步稳定的同质物种的热稳定性一致。这项研究表明，Env三聚体产生不同频率的有序与无序的异常三聚体，即使他们是遗传相同的。通过阴性选择天然样有序三聚体，我们建立了一种新的方法来获得衍生自亚型B和C的可溶性Env模拟物，以扩大用作候选疫苗免疫原。HIV包膜糖蛋白（Env）是病毒表面上唯一的病毒编码基因产物，因此是中和抗体的唯一靶标。广泛有效的HIV疫苗可能需要针对可变Env的保守元件产生稳健的中和抗体应答。对于成功的基于抗体的疫苗，可能需要HIV刺突的可溶性模拟物来产生能够中和多种HIV分离株的高滴度抗Env抗体。由于Env的全局序列多样性，产生这些可溶性刺突的多样性阵列将有益于设计用于科普这种病毒多样性的免疫策略。在这里，我们报告了一种新的纯化策略，然后对来自感染突出亚型B和C的两种可溶性HIV刺突进行了全面表征。我们证明，这些均匀的可溶性三聚体是忠实的模仿艾滋病毒穗中和抗体结合，电子显微镜和其他生物物理评估。拥有来自不同毒株的HIV刺突的可溶性和稳定的模拟物，既提高了我们对HIV刺突结构的认识，也扩大了潜在候选疫苗的亚型覆盖范围。
The structure of BG505 gp140 SOSIP, a soluble mimic of the native HIV-1 envelope glycoprotein (Env), marks the beginning of new era in Env structure-based immunogen design. Displaying a well-ordered quaternary structure, these subtype A-derived trimers display an excellent antigenic profile, discriminating recognition by broadly neutralizing antibodies (bNAbs) from non-broadly neutralizing antibodies (non-bNAbs), and provide a solid Env-based immunogenic platform starting point. Even with this important advance, obtaining homogeneous well-ordered soluble SOSIP trimers derived from other subtypes remains challenging. Here, we report the “rescue” of homogeneous well-ordered subtype B and C SOSIP trimers from a heterogeneous Env mixture using CD4 binding site-directed (CD4bs) non-bNAbs in a negative-selection purification process. These non-bNAbs recognize the primary receptor CD4bs only on disordered trimers but not on the native Env spike or well-ordered soluble trimers due to steric hindrance. Following negative selection to remove disordered oligomers, we demonstrated recovery of well-ordered, homogeneous trimers by electron microscopy (EM). We obtained 3D EM reconstructions of unliganded trimers, as well as in complex with sCD4, a panel of CD4bs-directed bNAbs, and the cleavage-dependent, trimer-specific bNAb, PGT151. Using bio-layer light interferometry (BLI) we demonstrated that the well-ordered trimers were efficiently recognized by bNAbs and poorly recognized by non-bNAbs, representing soluble mimics of the native viral spike. Biophysical characterization was consistent with the thermostability of a homogeneous species that could be further stabilized by specific bNAbs. This study revealed that Env trimers generate different frequencies of well-ordered versus disordered aberrant trimers even when they are genetically identical. By negatively selecting the native-like well-ordered trimers, we establish a new means to obtain soluble Env mimetics derived from subtypes B and C for expanded use as candidate vaccine immunogens. The HIV envelope glycoprotein (Env) is the sole virally encoded gene product on the surface of the virus and, as such, is the only target of neutralizing antibodies. A broadly efficacious HIV vaccine will likely need to generate a robust neutralizing antibody response directed at conserved elements of the variable Env. For a successful antibody-based vaccine, a soluble mimic of the HIV spike will likely be required to generate high-titer anti-Env antibodies capable of neutralizing a wide array of HIV isolates. Due to the global sequence diversity of Env, generating a diverse array of these soluble spikes will benefit immunization strategies designed to cope with such viral diversity. Here, we report a novel purification strategy followed by a comprehensive characterization of two soluble HIV spikes from infection-prominent subtypes, B and C. We demonstrate that these homogeneous soluble trimers are faithful mimics of the HIV spike by neutralizing antibody binding, electron microscopy and other biophysical assessments. Possessing soluble and stable mimics of the HIV spike derived from diverse strains improves both our knowledge of HIV spike architecture as shown here and extends subtype coverage of potential vaccine candidates.
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影响因子：
64.8
作者：
Huang J;Ofek G;Laub L;Louder MK;Doria-Rose NA;Longo NS;Imamichi H;Bailer RT;Chakrabarti B;Sharma SK;Alam SM;Wang T;Yang Y;Zhang B;Migueles SA;Wyatt R;Haynes BF;Kwong PD;Mascola JR;Connors M
通讯作者：
Connors M
影响因子：
5.5
作者：
Jones, Norman G.;DeCamp, Allan;Cao, Huyen
通讯作者：
Cao, Huyen
HIV-1 疫苗功效试验的免疫相关分析。
DOI：
10.1056/nejmoa1113425
发表时间：
2012-04-05
期刊：
The New England journal of medicine
影响因子：
--
作者：
Haynes BF;Gilbert PB;McElrath MJ;Zolla-Pazner S;Tomaras GD;Alam SM;Evans DT;Montefiori DC;Karnasuta C;Sutthent R;Liao HX;DeVico AL;Lewis GK;Williams C;Pinter A;Fong Y;Janes H;DeCamp A;Huang Y;Rao M;Billings E;Karasavvas N;Robb ML;Ngauy V;de Souza MS;Paris R;Ferrari G;Bailer RT;Soderberg KA;Andrews C;Berman PW;Frahm N;De Rosa SC;Alpert MD;Yates NL;Shen X;Koup RA;Pitisuttithum P;Kaewkungwal J;Nitayaphan S;Rerks-Ngarm S;Michael NL;Kim JH
通讯作者：
Kim JH
影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1073/pnas.1103012108
发表时间：
2011-07-05
影响因子：
11.1
作者：
Burton, Dennis R.;Hessell, Ann J.;Moore, John P.
通讯作者：
Moore, John P.