Human immunodeficiency virus type-1 (HIV-1) antigenic variation poses a great challenge for vaccine immunogen design to elicit broadly neutralizing antibodies (bNAbs). Over the last 10–15 years, great progress has been made to understand the conserved sites of sensitivity on HIV envelope glycoprotein spikes targeted by bNAbs. Plasma neutralization mapping and monoclonal antibody isolation efforts have revealed five major conserved epitope clusters. Most of this work has focused on subtype B and C-infected Caucasian or African donors. It is not clear if the same epitopes and epitope rank order preferences are also true in donors infected with different HIV-1 subtypes and with different racial backgrounds. To investigate this point, in this study we report the first attempt to profile the bNAb specificities of CRF01_AE-infected Malaysian plasmas. We first measured neutralization titers of 21 plasmas against a subtype A, B, and AE pseudovirus panel. This revealed that 14% (3 of 21) plasmas had cross-clade breadth. Focusing on the cross-neutralizing plasma P9, we used AE and JR-FL mutant pseudoviruses, gp120 monomer interference, and native polyacrylamide gel electrophoresis to better understand the neutralization specificity. P9 demonstrates CD4-binding-site specificity with trimer dependence and D368 independence.

中文翻译：

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来自马来西亚的交叉中和 CRF01_AE 感染血浆靶向人类免疫缺陷病毒 1 型包膜糖蛋白的 CD4 结合位点

人类免疫缺陷病毒 1 型 (HIV-1) 抗原变异对疫苗免疫原设计以引发广泛中和抗体 (bNAb) 提出了巨大挑战。在过去的 10-15 年中，在了解 bNAb 靶向的 HIV 包膜糖蛋白尖峰的敏感性保守位点方面取得了很大进展。血浆中和作图和单克隆抗体分离工作揭示了五个主要的保守表位簇。这项工作的大部分集中在 B 和 C 亚型感染的高加索或非洲供体上。目前尚不清楚相同的表位和表位排序偏好是否也适用于感染不同 HIV-1 亚型和不同种族背景的供体。为了研究这一点，在本研究中，我们首次尝试分析 CRF01_AE 感染的马来西亚血浆的 bNAb 特异性。我们首先测量了 21 种血浆对 A、B 和 AE 假病毒组的中和滴度。这表明 14%（21 个中的 3 个）血浆具有跨进化枝宽度。针对交叉中和血浆 P9，我们使用 AE 和 JR-FL 突变假病毒、gp120 单体干扰和天然聚丙烯酰胺凝胶电泳来更好地了解中和特异性。P9 展示了具有三聚体依赖性和 D368 独立性的 CD4 结合位点特异性。