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SARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates.
Emerging Microbes & Infections ( IF 8.4 ) Pub Date : 2020-09-24 , DOI: 10.1080/22221751.2020.1821583
Tingting Li 1, 2 , Qingbing Zheng 1, 2 , Hai Yu 1, 2 , Dinghui Wu 3 , Wenhui Xue 1, 2 , Hualong Xiong 1, 2 , Xiaofen Huang 1, 2 , Meifeng Nie 1, 2 , Mingxi Yue 1, 2 , Rui Rong 1, 2 , Sibo Zhang 1, 2 , Yuyun Zhang 1, 2 , Yangtao Wu 1, 2 , Shaojuan Wang 1, 2 , Zhenghui Zha 1, 2 , Tingting Chen 1, 2 , Tingting Deng 1, 2 , Yingbin Wang 1, 2 , Tianying Zhang 1, 2 , Yixin Chen 1, 2 , Quan Yuan 1, 2 , Qinjian Zhao 1, 2 , Jun Zhang 1, 2 , Ying Gu 1, 2 , Shaowei Li 1, 2 , Ningshao Xia 1, 2, 4
Affiliation  

The current coronavirus disease 2019 (COVID-19) pandemic was the result of the rapid transmission of a highly pathogenic coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for which there is no efficacious vaccine or therapeutic. Toward the development of a vaccine, here we expressed and evaluated as potential candidates four versions of the spike (S) protein using an insect cell expression system: receptor binding domain (RBD), S1 subunit, the wild-type S ectodomain (S-WT), and the prefusion trimer-stabilized form (S-2P). We showed that RBD appears as a monomer in solution, whereas S1, S-WT, and S-2P associate as homotrimers with substantial glycosylation. Cryo-electron microscopy analyses suggested that S-2P assumes an identical trimer conformation as the similarly engineered S protein expressed in 293 mammalian cells but with reduced glycosylation. Overall, the four proteins confer excellent antigenicity with convalescent COVID-19 patient sera in enzyme-linked immunosorbent assay (ELISA), yet show distinct reactivities in immunoblotting. RBD, S-WT and S-2P, but not S1, induce high neutralization titres (>3-log) in mice after a three-round immunization regimen. The high immunogenicity of S-2P could be maintained at the lowest dose (1 μg) with the inclusion of an aluminium adjuvant. Higher doses (20 μg) of S-2P can elicit high neutralization titres in non-human primates that exceed 40-times the mean titres measured in convalescent COVID-19 subjects. Our results suggest that the prefusion trimer-stabilized SARS-CoV-2 S-protein from insect cells may offer a potential candidate strategy for the development of a recombinant COVID-19 vaccine.



中文翻译:

昆虫细胞中产生的SARS-CoV-2尖峰在非人类灵长类动物中引起很高的中和效价。

当前的2019年冠状病毒疾病(COVID-19)大流行是高致病性冠状病毒,严重的急性呼吸综合征冠状病毒2(SARS-CoV-2)快速传播的结果,目前尚无有效的疫苗或治疗剂。为了开发疫苗,在这里我们使用昆虫细胞表达系统表达并评估了穗蛋白(S)蛋白质的四个版本作为潜在候选者:受体结合域(RBD),S1亚基,野生型S胞外域(S- WT)和融合前三聚体稳定形式(S-2P)。我们显示,RBD在溶液中以单体形式出现,而S1,S-WT和S-2P以均三聚体的形式伴有大量糖基化。低温电子显微镜分析表明,S-2P假定的三聚体构象与在293个哺乳动物细胞中表达的类似工程化的S蛋白相同,但糖基化程度降低。总体而言,这四种蛋白质在酶联免疫吸附测定(ELISA)中与恢复期的COVID-19患者血清具有优异的抗原性,但在免疫印迹中却表现出不同的反应性。三轮免疫方案后,RBD,S-WT和S-2P而非S1在小鼠中诱导高中和效价(> 3-log)。含有铝佐剂可以将S-2P的高免疫原性维持在最低剂量(1μg)。较高剂量(20μg)的S-2P可以在非人灵长类动物中引起高中和滴度,其超过恢复期COVID-19受试者中平均滴度的40倍。

更新日期:2020-09-24
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