The recent SARS-CoV-2 pandemic has taught the world a costly lesson about the devastating consequences of viral disease outbreaks but also, the remarkable impact of vaccination in limiting life and economic losses. Vaccination against human Hepatitis B Virus (HBV), a major human pathogen affecting 290 million people worldwide, remains a key action towards viral hepatitis elimination by 2030. To meet this goal, the development of improved HBV antigens is critical to overcome non-responsiveness to standard vaccines based on the yeast-produced, small (S) envelope protein. We have recently shown that combining relevant immunogenic determinants of S and large (L) HBV proteins in chimeric antigens markedly enhances the anti-HBV immune response. However, the demand for cost-efficient, high-quality antigens remains challenging. This issue could be addressed by using plants as versatile and rapidly scalable protein production platforms. Moreover, the recent generation of plants lacking β-1,2-xylosyltransferase and α-1,3-fucosyltransferase activities (FX-KO), by CRISPR/Cas9 genome editing, enables production of proteins with “humanized” N-glycosylation. In this study, we investigated the impact of plant N-glycosylation on the immunogenic properties of a chimeric HBV S/L vaccine candidate produced in wild-type and FX-KO Nicotiana benthamiana. Prevention of β-1,2-xylose and α-1,3-fucose attachment to the HBV antigen significantly increased the immune response in mice, as compared with the wild-type plant-produced counterpart. Notably, the antibodies triggered by the FX-KO-made antigen neutralized more efficiently both wild-type HBV and a clinically relevant vaccine escape mutant. Our study validates in premiere the glyco-engineered Nicotiana benthamiana as a substantially improved host for plant production of glycoprotein vaccines.

中文翻译：

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CRISPR/Cas9 编辑的本塞姆氏烟草中的“人源化”N-糖基化途径显着增强了 S/preS1 乙型肝炎病毒抗原的免疫原性以及疫苗接种小鼠中的病毒中和抗体反应


最近的 SARS-CoV-2 大流行给世界上了一堂代价高昂的课：病毒性疾病爆发的破坏性后果，以及疫苗接种在限制生命和经济损失方面的显着影响。人类乙型肝炎病毒 (HBV) 是一种影响全球 2.9 亿人的主要人类病原体，它仍然是到 2030 年消除病毒性肝炎的关键行动。为了实现这一目标，开发改进的 HBV 抗原对于克服对乙型肝炎病毒无反应的情况至关重要。基于酵母产生的小 (S) 包膜蛋白的标准疫苗。我们最近发现，在嵌合抗原中结合 S 和大 (L) HBV 蛋白的相关免疫原性决定簇可显着增强抗 HBV 免疫反应。然而，对具有成本效益的高质量抗原的需求仍然具有挑战性。这个问题可以通过使用植物作为多功能且快速扩展的蛋白质生产平台来解决。此外，通过 CRISPR/Cas9 基因组编辑，最新一代缺乏 β-1,2-木糖基转移酶和 α-1,3-岩藻糖基转移酶活性 (FX-KO) 的植物能够生产具有“人源化”N-糖基化的蛋白质。在这项研究中，我们研究了植物 N-糖基化对野生型和 FX-KO烟草中生产的嵌合 HBV S/L 候选疫苗免疫原性的影响。与野生型植物产生的对应物相比，阻止 β-1,2-木糖和 α-1,3-岩藻糖与 HBV 抗原的附着可显着增强小鼠的免疫反应。值得注意的是，由 FX-KO 制造的抗原触发的抗体更有效地中和野生型 HBV 和临床相关的疫苗逃逸突变体。 我们的研究首次验证了糖基改造的本塞姆氏烟草作为植物生产糖蛋白疫苗的显着改良宿主。