Bispecific and Trispecific Anti-Env Antibodies for Eliminating HIV Reservoir Cells

项目摘要/摘要 抗体是对抗病毒感染的有力武器。它们可以中和病毒的传染性 颗粒以及通过抗体依赖等机制介导对受感染细胞的杀伤 细胞介导的细胞毒性(ADCC)。在过去的五年中，我们集团利用了在 抗体工程领域，以及越来越多的广谱中和抗体(BNAbs)，由 其他研究人员，构建新的双特异性抗体，具有精致的抗病毒广度和效力 预防HIV-1的目的。现在，通过这项提议，我们希望将我们的抗体工程努力扩大到 产生一组双特异性和三特异性抗体，这些抗体是针对杀死Env表达而优化的 细胞，以及重要的性质，如药代动力学。而不是筛查病毒中和 正如我们之前所做的那样，我们现在将设计和筛选一个针对环境的多特定 体外细胞结合和细胞杀伤活性的抗体。性能最好的抗体构建体将 在人源化的小鼠模型中系统地评估其对细胞杀伤能力的影响， 它们在激活后限制或消除潜伏的储集层细胞的能力，以及它们防止或限制 HIV-1潜伏库的建立。随着我们对HIV-1bNAbs和抗体的了解 工程学，我们对病毒动力学的深刻理解和我们以前研究HIV-1潜伏期的经验 将使储集层对实验的设计、实施和解释产生影响，以评估和 量化我们的顶级抗体构建物在人源化小鼠中的抗病毒效果。 我们的团队已经成功地设计出两种具有精致的HIV-1中和活性的双特异性抗体，以及 自那以后，我们将我们在抗体工程方面的技术诀窍扩展到构建双特异性或三特异性 以Env为靶标的抗体，目的是促进清除受感染的细胞。最后，我们希望 向现场提供一到两种可用于消除潜伏的多特异性抗体 将储存细胞作为多管齐下根除艾滋病毒-1方法的一个关键组成部分。

PROJECT SUMMARY/ABSTRACT Antibodies constitute a powerful weapon to fight viral infections. They could neutralize the infectivity of virus particles as well as mediate killing of productively infected cells via mechanisms such as antibody-dependent cell-mediated cytotoxicity (ADCC). In the past five years, our group has utilized the advances made in the antibody engineering field, as well as the growing lists of broadly neutralization antibodies (bNAbs) identified by other investigators, to construct new bispecific antibodies that have exquisite antiviral breadth and potency for the purpose of HIV-1 prevention. Now, with this proposal, we wish to expand our antibody engineering effort to generate a collection of bispecific and trispecific antibodies that are optimized for killing of Env-expressing cells, and for important properties such as pharmacokinetics. Instead of screening for virus-neutralization activity as we have previously done, we will now engineer and screen a library of Env-targeting multi-specific antibodies for cell-binding and cell-killing activities in vitro. The best performing antibody constructs will then be evaluated systematically in vivo in a humanized mouse model for the effect on their cell-killing capacity, their ability to restrict or eliminate latent reservoir cells after activation, and their ability to prevent or limit the establishment of the HIV-1 latent reservoir. Along with our knowledge of HIV-1 bNAbs and antibody engineering, our deep understanding of viral dynamics and our prior experience studying the HIV-1 latent reservoir will be brought to bear on the design, conduct, and interpretation of experiments to evaluate and quantify the antiviral effects of our top antibody constructs in humanized mice. Our group has successfully engineered two bispecific antibodies with exquisite HIV-1-neutralizing activity, and we have since extended our know-how in antibody engineering to the construction of bispecific or trispecific antibodies that target Env for the purpose of facilitating the elimination of infected cells. In the end, we hope to offer to the field one or two multi-specific antibodies that could be applied toward the elimination of latent reservoir cells as one critical component of a multi-pronged approach to HIV-1 eradication.