Radford Caelan E, Schommers Philipp, Gieselmann Lutz, Crawford Katharine H D, Dadonaite Bernadeta, Yu Timothy C, Dingens Adam S, Overbaugh Julie, Klein Florian, Bloom Jesse D
Molecular and Cellular Biology Graduate Program, University of Washington, and Basic Sciences Division, Fred Hutch Cancer Center, Seattle, Washington, 98109, USA.
Basic Sciences Division and Computational Biology Program, Fred Hutchinson Cancer Center, Seattle, Washington, 98109, USA.
bioRxiv. 2023 Mar 24:2023.03.23.533993. doi: 10.1101/2023.03.23.533993.
Understanding the specificities of human serum antibodies that broadly neutralize HIV can inform prevention and treatment strategies. Here we describe a deep mutational scanning system that can measure the effects of combinations of mutations to HIV envelope (Env) on neutralization by antibodies and polyclonal serum. We first show that this system can accurately map how all functionally tolerated mutations to Env affect neutralization by monoclonal antibodies. We then comprehensively map Env mutations that affect neutralization by a set of human polyclonal sera known to target the CD4-binding site that neutralize diverse strains of HIV. The neutralizing activities of these sera target different epitopes, with most sera having specificities reminiscent of individual characterized monoclonal antibodies, but one sera targeting two epitopes within the CD4 binding site. Mapping the specificity of the neutralizing activity in polyclonal human serum will aid in assessing anti-HIV immune responses to inform prevention strategies.
了解能广泛中和HIV的人血清抗体的特异性可为预防和治疗策略提供依据。在此，我们描述了一种深度突变扫描系统，该系统可测量HIV包膜（Env）突变组合对抗体和多克隆血清中和作用的影响。我们首先表明，该系统能够准确绘制出所有对Env功能耐受的突变如何影响单克隆抗体的中和作用。然后，我们全面绘制了Env突变，这些突变会影响一组已知靶向CD4结合位点以中和多种HIV毒株的人多克隆血清的中和作用。这些血清的中和活性靶向不同的表位，大多数血清具有类似于单个已表征单克隆抗体的特异性，但有一种血清靶向CD4结合位点内的两个表位。绘制多克隆人血清中中和活性的特异性将有助于评估抗HIV免疫反应，为预防策略提供信息。
