Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses.
Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses.
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DOI：
10.1371/journal.pone.0119608
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Berman PW
中科院分区：
文献类型：
--
作者：
O'Rourke SM;Sutthent R;Phung P;Mesa KA;Frigon NL;To B;Horthongkham N;Limoli K;Wrin T;Berman PW
了解对中和抗体的敏感性和抵抗力的分子决定因素，对于开发旨在预防艾滋病毒感染的疫苗至关重要。在这项研究中，我们使用遗传方法来表征HIV包膜蛋白中自然发生的多态，这些多态赋予中和敏感性或抵抗力。从CRF01_AE病毒感染的个体中构建了来源于病毒准种的紧密相关的囊膜基因文库。用含有广谱中和抗体的血浆筛选文库，并选择中和敏感和抗性突变体进行序列分析。体外诱变使我们能够在三个个体中鉴定出对这些抗体中和产生抵抗的单一氨基酸变化。这三个突变都在四链V1/V2结构域β-Sheet结构中A链C端和B链N端之间的高变量连接肽附近产生了N-连接糖基化位点(两个位于N136，一个位于N149)。尽管N136以前与广谱中和单抗的结合有关，但这个糖基化位点似乎抑制了HIV-1感染者血浆中中和抗体的结合。以往的研究报道，与慢性感染分离的病毒相比，传播型方正病毒的V1/V2结构域长度更短，糖基化位点更少。我们的结果表明，基于CRF01_AE病毒重组包膜蛋白的疫苗免疫原可以通过包含缺乏这些糖基化位点的包膜蛋白来改进。这一策略可能会提高在部分成功的RV144 HIV疫苗试验中使用的疫苗的效力，在该试验中，两种CRF01_AE免疫原(来自A244和TH023分离株)都具有N136和N149的糖基化位点。
Understanding the molecular determinants of sensitivity and resistance to neutralizing antibodies is critical for the development of vaccines designed to prevent HIV infection. In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. In vitro mutagenesis allowed us to identify single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies. All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149.
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DOI：
10.1093/bioinformatics/btq224
发表时间：
2010-08-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Katoh K;Toh H
通讯作者：
Toh H
影响因子：
82.9
作者：
通讯作者：
--
影响因子：
6.4
作者：
Gianella, Sara;Richman, Douglas D.
通讯作者：
Richman, Douglas D.
影响因子：
56.9
作者：
BURTON, DR;PYATI, J;BARBAS, CF
通讯作者：
BARBAS, CF
影响因子：
3.7
作者：
Berman, PW;Huang, W;Gregory, TJ
通讯作者：
Gregory, TJ