Protein Engineering and Therapeutics Group, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, 20132, Milan, Italy.
INSERM, UMRS-839, Institut du Fer à Moulin, 75005, Paris, France.
Sci Rep. 2018 Jan 30;8(1):1890. doi: 10.1038/s41598-018-20300-9.
Efforts to improve existing anti-HIV-1 therapies or develop preventatives have identified CCR5 as an important target and CCL5 as an ideal scaffold to sculpt potent HIV-1 entry inhibitors. We created novel human CCL5 variants that exhibit exceptional anti-HIV-1 features using recombinant lactobacilli (exploited for live microbicide development) as a screening platform. Protein design, expression and anti-HIV-1 activity flowed in iterative cycles, with a stepwise integration of successful mutations and refinement of an initial CCL5 mutant battery towards the generation of two ultimate CCL5 derivatives, a CCR5 agonist and a CCR5 antagonist with similar anti-HIV-1 potency. The CCR5 antagonist was tested in human macrophages and against primary R5 HIV-1 strains, exhibiting cross-clade low picomolar IC activity. Moreover, its successful combination with several HIV-1 inhibitors provided the ground for conceiving therapeutic and preventative anti-HIV-1 cocktails. Beyond HIV-1 infection, these CCL5 derivatives may now be tested against several inflammation-related pathologies where the CCL5:CCR5 axis plays a relevant role.
为了改进现有的抗 HIV-1 疗法或开发预防药物，研究人员已经将 CCR5 确定为一个重要的靶点，将 CCL5 确定为构建高效 HIV-1 进入抑制剂的理想支架。我们使用重组乳杆菌（用于开发活体杀菌剂）作为筛选平台，创造了具有优异抗 HIV-1 特性的新型人类 CCL5 变体。蛋白质设计、表达和抗 HIV-1 活性在迭代循环中进行，成功的突变逐步整合，并对初始 CCL5 突变体电池进行改进，最终生成两种 CCL5 衍生物，一种是 CCR5 激动剂，一种是 CCR5 拮抗剂，具有相似的抗 HIV-1 效力。该 CCR5 拮抗剂在人巨噬细胞和原发性 R5 HIV-1 株中进行了测试，表现出跨谱系低皮摩尔 IC 活性。此外，它与几种 HIV-1 抑制剂的成功结合为构思治疗和预防 HIV-1 的鸡尾酒疗法提供了基础。除了 HIV-1 感染，这些 CCL5 衍生物现在可能会在几种与炎症相关的病理中进行测试，其中 CCL5:CCR5 轴发挥着重要作用。
