Fan Chengcheng, Keeffe Jennifer R, Malecek Kathryn E, Cohen Alexander A, West Anthony P, Baharani Viren A, Rorick Annie V, Gao Han, Gnanapragasam Priyanthi N P, Rho Semi, Alvarez Jaasiel, Segovia Luisa N, Hatziioannou Theodora, Bieniasz Paul D, Bjorkman Pamela J
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125.
Laboratory of Retrovirology, The Rockefeller University, New York, NY 10065.
Proc Natl Acad Sci U S A. 2025 May 27;122(21):e2501637122. doi: 10.1073/pnas.2501637122. Epub 2025 May 22.
Broad immune responses are needed to mitigate viral evolution and escape. To induce antibodies against conserved receptor-binding domain (RBD) regions of SARS-like betacoronavirus (sarbecovirus) spike proteins that recognize SARS-CoV-2 variants of concern and zoonotic sarbecoviruses, we developed mosaic-8b RBD nanoparticles presenting eight sarbecovirus RBDs arranged randomly on a 60-mer nanoparticle. Mosaic-8b immunizations protected animals from challenges from viruses whose RBDs were matched or mismatched to those on nanoparticles. Here, we describe neutralizing mAbs isolated from mosaic-8b-immunized rabbits, some on par with Pemgarda, the only currently FDA-approved therapeutic mAb. Deep mutational scanning, in vitro selection of spike resistance mutations, and single-particle cryo-electron microscopy structures of spike-antibody complexes demonstrated targeting of conserved RBD epitopes. Rabbit mAbs included critical D-gene segment RBD-recognizing features in common with human anti-RBD mAbs, despite rabbit genomes lacking an equivalent human D-gene segment, thus demonstrating that the immune systems of humans and other mammals can utilize different antibody gene segments to arrive at similar modes of antigen recognition. These results suggest that animal models can be used to elicit anti-RBD mAbs with similar properties to those raised in humans, which can then be humanized for therapeutic use, and that mosaic RBD nanoparticle immunization coupled with multiplexed screening represents an efficient way to generate and select broadly cross-reactive therapeutic pan-sarbecovirus and pan-SARS-CoV-2 variant mAbs.
需要广泛的免疫反应来减轻病毒进化和逃逸。为了诱导针对识别严重急性呼吸综合征冠状病毒2(SARS-CoV-2)关注变体和人畜共患沙贝病毒的SARS样β冠状病毒(沙贝病毒)刺突蛋白保守受体结合域(RBD)区域的抗体,我们开发了镶嵌8b RBD纳米颗粒,其在一个60聚体纳米颗粒上随机排列有8个沙贝病毒RBD。镶嵌8b免疫可保护动物免受RBD与纳米颗粒上的RBD匹配或不匹配的病毒的攻击。在此,我们描述了从经镶嵌8b免疫的兔子中分离出的中和单克隆抗体(mAb),其中一些与目前唯一获得美国食品药品监督管理局(FDA)批准的治疗性mAb Pemgarda相当。深度突变扫描、刺突抗性突变的体外选择以及刺突-抗体复合物的单颗粒冷冻电子显微镜结构表明,这些抗体靶向保守的RBD表位。兔源mAb包括与人类抗RBD mAb共有的关键D基因片段RBD识别特征,尽管兔基因组缺乏等效的人类D基因片段,这表明人类和其他哺乳动物的免疫系统可以利用不同的抗体基因片段来实现相似的抗原识别模式。这些结果表明,动物模型可用于引发具有与人类产生的抗体相似特性的抗RBD mAb,然后可将其人源化用于治疗,并且镶嵌RBD纳米颗粒免疫与多重筛选相结合是生成和选择具有广泛交叉反应性的治疗性泛沙贝病毒和泛SARS-CoV-2变体mAb的有效方法。
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