Torrey Pines Institute for Molecular Studies, San Diego, California, USA.
J Virol. 2012 Apr;86(7):3574-87. doi: 10.1128/JVI.06938-11. Epub 2012 Feb 1.
Hypothetically, since native HIV-1 Env trimers are exclusively recognized by neutralizing antibodies, they might induce the neutralizing antibodies in a vaccine setting. This idea has not been evaluated due to the difficulty of separating trimers from nonfunctional Env (uncleaved gp160 and gp41 stumps). The latter are immunodominant and induce nonneutralizing antibodies. We previously showed that nonfunctional Env can be selectively cleared from virus-like particle (VLP) surfaces by enzyme digests (E. T. Crooks, T. Tong(,) K. Osawa, and J. M. Binley, J.Virol. 85:5825, 2011). Here, we investigated the effects of these digests on the antigenicity of VLPs and their sensitivity to neutralization. Before digestion, WT VLPs (bearing wild-type Env) and UNC VLPs (bearing uncleaved gp160) were recognized by various Env-specific monoclonal antibodies (MAbs), irrespective of their neutralizing activity, a result which is consistent with the presence of nonfunctional Env. After digestion, only neutralizing MAbs recognized WT VLPs, consistent with selective removal of nonfunctional Env (i.e., "trimer VLPs"). Digests eliminated the binding of all MAbs to UNC VLPs, again consistent with removal of nonfunctional Env. An exception was MAb 2F5, which weakly bound to digested UNC VLPs and bald VLPs (bearing no Env), perhaps due to lipid cross-reactivity. Trimer VLPs were infectious, and their neutralization sensitivity was largely comparable to that of undigested WT VLPs. However, they were ∼100-fold more sensitive to the MAbs 4E10 and Z13e1, suggesting increased exposure of the gp41 base. Importantly, a scatterplot analysis revealed a strong correlation between MAb binding and neutralization of trimer VLPs. This suggests that trimer VLPs bear essentially pure native trimer that should allow its unfettered evaluation in a vaccine setting.
假设天然 HIV-1 Env 三聚体仅被中和抗体识别,那么它们可能在疫苗接种环境中诱导中和抗体。由于难以将三聚体与无功能的 Env(未切割的 gp160 和 gp41 残基)分离,因此尚未评估此想法。后者是免疫显性的,会诱导非中和抗体。我们之前表明,可以通过酶消化选择性地从病毒样颗粒(VLP)表面清除无功能的 Env(E. T. Crooks、T. Tong、K. Osawa 和 J. M. Binley,J.Virol. 85:5825,2011)。在这里,我们研究了这些消化物对 VLP 抗原性及其对中和的敏感性的影响。在消化之前,WT VLP(携带野生型 Env)和 UNC VLP(携带未切割的 gp160)被各种 Env 特异性单克隆抗体(MAb)识别,无论其中和活性如何,这一结果与存在无功能的 Env 一致。消化后,只有中和 MAb 识别 WT VLP,这与选择性去除无功能的 Env(即“三聚体 VLP”)一致。消化物消除了所有 MAb 与 UNC VLP 的结合,这再次与无功能 Env 的去除一致。一个例外是 MAb 2F5,它与消化的 UNC VLP 和无毛 VLP(不携带 Env)弱结合,可能是由于脂质交叉反应。三聚体 VLP 具有感染性,其中和敏感性与未消化的 WT VLP 基本相当。然而,它们对 MAb 4E10 和 Z13e1 的敏感性提高了约 100 倍,表明 gp41 基部的暴露增加。重要的是,散点图分析显示 MAb 结合与三聚体 VLP 的中和之间存在很强的相关性。这表明三聚体 VLP 携带基本上是纯的天然三聚体,这应该允许在疫苗接种环境中不受限制地评估它。