CD4-based decoy approaches against HIV-1 are attractive options for long-term viral control, but initial designs, including soluble CD4 (sCD4) and CD4-Ig, were ineffective. To evaluate a therapeutic that more accurately mimics HIV-1 target cells compared with monomeric sCD4 and dimeric CD4-Ig, we generated virus-like nanoparticles that present clusters of membrane-associated CD4 (CD4-VLPs) to permit high-avidity binding of trimeric HIV-1 envelope spikes. In neutralization assays, CD4-VLPs were >12,000-fold more potent than sCD4 and CD4-Ig and >100-fold more potent than the broadly neutralizing antibody (bNAb) 3BNC117, with >12,000-fold improvements against strains poorly neutralized by 3BNC117. CD4-VLPs also neutralized patient-derived viral isolates that were resistant to 3BNC117 and other bNAbs. Intraperitoneal injections of CD4-CCR5-VLP produced only subneutralizing plasma concentrations in HIV-1–infected humanized mice but elicited CD4-binding site mutations that reduced viral fitness. All mutant viruses showed reduced sensitivity to sCD4 and CD4-Ig but remained sensitive to neutralization by CD4-VLPs in vitro. In vitro evolution studies demonstrated that CD4-VLPs effectively controlled HIV-1 replication at neutralizing concentrations, and viral escape was not observed. Moreover, CD4-VLPs potently neutralized viral swarms that were completely resistant to CD4-Ig, suggesting that escape pathways that confer resistance against conventional CD4-based inhibitors are ineffective against CD4-VLPs. These findings suggest that therapeutics that mimic HIV-1 target cells could prevent viral escape by exposing a universal vulnerability of HIV-1: the requirement to bind CD4 on a target cell. We propose that therapeutic and delivery strategies that ensure durable bioavailability need to be developed to translate this concept into a clinically feasible functional cure therapy.

基于CD4的针对HIV-1的诱饵方法是长期病毒控制的诱人选择，但包括可溶性CD4（sCD4）和CD4-Ig在内的初始设计均无效。为了评估与单体sCD4和二聚体CD4-Ig相比更精确地模拟HIV-1靶细胞的治疗剂，我们生成了病毒样纳米颗粒，它们呈现与膜相关的CD4（CD4-VLPs）簇，可高度结合三聚体HIV-1包膜尖峰。在中和试验中，CD4-VLPs的效力比sCD4和CD4-Ig高12,000倍，效力比广泛中和抗体（bNAb）3BNC117高100倍，对3BNC117中和效果较差的菌株的效力提高了12,000倍。CD4-VLP还中和了对3BNC117和其他bNAb耐药的患者源性病毒分离株。腹膜内注射CD4-CCR5-VLP在感染HIV-1的人源化小鼠中仅产生低于中和的血浆浓度，但引起CD4结合位点突变，降低了病毒适应性。所有突变病毒在体外对sCD4和CD4-Ig的敏感性均降低，但对CD4-VLP的中和作用仍然敏感。体外进化研究表明，CD4-VLP在中和浓度下可有效控制HIV-1复制，并且未观察到病毒逃逸。此外，CD4-VLP有效地中和了对CD4-Ig完全耐药的病毒群，表明赋予对常规CD4抑制剂抗性的逃逸途径对CD4-VLP无效。这些发现表明，模仿HIV-1目标细胞的疗法可通过暴露HIV-1的普遍脆弱性来防止病毒逃逸：在靶细胞上结合CD4的要求。我们建议需要开发确保持久生物利用度的治疗和给药策略，以将这一概念转化为临床上可行的功能性治疗方法。