Umotoy Jeffrey C, Kroon Pascal Z, Man Shirley, van Dort Karel A, Atabey Tugba, Schriek Angela I, Dekkers Gillian, Herrera-Carrillo Elena, Geijtenbeek Teunis B H, Heukers Raimond, Kootstra Neeltje A, van Gils Marit J, de Taeye Steven W
Department of Medical Microbiology and Infection Prevention, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, the Netherlands.
Amsterdam Institute for Immunology and Infectious Diseases, Amsterdam, the Netherlands.
iScience. 2024 Sep 13;27(10):110958. doi: 10.1016/j.isci.2024.110958. eCollection 2024 Oct 18.
HIV-1 alters the dynamics and distribution of tetraspanins, a group of proteins integral to membrane organization, to facilitate both entry and egress. Notably, the tetraspanin CD9 is dysregulated during HIV-1 infection, correlating with multifaceted effects on viral replication. Here, we generated llama-derived nanobodies against CD9 to restrict HIV-1 replication. We immunized llamas with recombinant large extracellular loop of CD9 and identified eight clonally distinct nanobodies targeting CD9, each exhibiting a range of affinities and differential binding to cell surface-expressed CD9. Notably, nanobodies T2C001 and T2C002 demonstrated low nanomolar affinities and exhibited differential sensitivities against endogenous and overexpressed CD9 on the cell surface. Although CD9-directed nanobodies did not impede the early stages of HIV-1 life cycle, they effectively inhibited virus-induced syncytia formation and virus replication in T cells and monocyte-derived macrophages. This discovery opens new avenues for host-targeted therapeutic strategies, potentially augmenting existing antiretroviral treatments for HIV-1.
HIV-1会改变四跨膜蛋白的动态变化和分布，这类蛋白是膜组织不可或缺的一部分，HIV-1借此促进病毒的进入和释放。值得注意的是，在HIV-1感染过程中，四跨膜蛋白CD9的表达失调，这与对病毒复制的多方面影响相关。在此，我们制备了针对CD9的羊驼源纳米抗体，以限制HIV-1的复制。我们用重组的CD9大细胞外环免疫羊驼，并鉴定出8种针对CD9的克隆不同的纳米抗体，每种纳米抗体都表现出一系列亲和力以及与细胞表面表达的CD9的差异结合。值得注意的是，纳米抗体T2C001和T2C002表现出低纳摩尔亲和力，并对细胞表面内源性和过表达的CD9表现出不同的敏感性。虽然针对CD9的纳米抗体并不妨碍HIV-1生命周期的早期阶段，但它们能有效抑制T细胞和单核细胞来源的巨噬细胞中病毒诱导的合胞体形成和病毒复制。这一发现为以宿主为靶点的治疗策略开辟了新途径，有可能增强现有的HIV-1抗逆转录病毒治疗方法。
