Yang Ming, Sun Haiyan, Lai Huafang, Neupane Biswas, Bai Fengwei, Steinkellner Herta, Chen Qiang
The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ 85225, USA.
Department of Cell and Molecular Biology, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
Vaccines (Basel). 2023 Mar 29;11(4):755. doi: 10.3390/vaccines11040755.
Monoclonal antibodies (mAb) against the envelope (E) protein of Zika virus (ZIKV) have shown great potential as therapeutics against the Zika epidemics. However, their use as a therapy may predispose treated individuals to severe infection by the related dengue virus (DENV) via antibody-dependent enhancement of infection (ADE). Here, we generated a broadly neutralizing flavivirus mAb, ZV1, with an identical protein backbone but different Fc glycosylation profiles. The three glycovariants, produced in wild-type (WT) and glycoengineered ΔXF plants and in Chinese hamster ovary cells (ZV1, ZV1, and ZV1), respectively, showed equivalent neutralization potency against both ZIKV and DENV. By contrast, the three mAb glycoforms demonstrated drastically different ADE activity for DENV and ZIKV infection. While ZV1 and ZV1 showed ADE activity upon DENV and ZIKV infection, ZV1 totally forwent its ADE. Importantly, all three glycovariants exhibited antibody-dependent cellular cytotoxicity (ADCC) against virus-infected cells, with increased potency by the fucose-free ZV1 glycoform. Moreover, the in vivo efficacy of the ADE-free ZV1 was demonstrated in a murine model. Collectively, we demonstrated the feasibility of modulating ADE by Fc glycosylation, thereby establishing a novel approach for improving the safety of flavivirus therapeutics. Our study also underscores the versatile use of plants for the rapid expression of complex human proteins to reveal novel insight into antibody function and viral pathogenesis.
针对寨卡病毒（ZIKV）包膜（E）蛋白的单克隆抗体（mAb）已显示出作为对抗寨卡疫情疗法的巨大潜力。然而，将其用作治疗方法可能会使接受治疗的个体通过抗体依赖性感染增强（ADE）而更容易受到相关登革病毒（DENV）的严重感染。在此，我们生成了一种具有相同蛋白质骨架但不同Fc糖基化谱的广谱中和黄病毒单克隆抗体ZV1。分别在野生型（WT）和糖基工程化的ΔXF植物以及中国仓鼠卵巢细胞中产生的三种糖变体（ZV1、ZV1和ZV1）对ZIKV和DENV均显示出等效的中和效力。相比之下，这三种单克隆抗体糖型对DENV和ZIKV感染表现出截然不同的ADE活性。虽然ZV1和ZV1在DENV和ZIKV感染时表现出ADE活性，但ZV1完全没有ADE活性。重要的是，所有三种糖变体均对病毒感染细胞表现出抗体依赖性细胞毒性（ADCC），无岩藻糖的ZV1糖型的效力增强。此外，在小鼠模型中证明了无ADE的ZV1的体内疗效。总体而言，我们证明了通过Fc糖基化调节ADE的可行性，从而建立了一种提高黄病毒治疗安全性的新方法。我们的研究还强调了植物在快速表达复杂人类蛋白质以揭示抗体功能和病毒发病机制新见解方面的广泛用途。
