SARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates.
SARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates.
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昆虫细胞中产生的 SARS-CoV-2 刺突在非人类灵长类动物中引起高中和滴度
DOI：
10.1080/22221751.2020.1821583
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发表时间：
2020-12
影响因子：
13.2
通讯作者：
Xia N
中科院分区：
文献类型：
--
作者：
Li T;Zheng Q;Yu H;Wu D;Xue W;Xiong H;Huang X;Nie M;Yue M;Rong R;Zhang S;Zhang Y;Wu Y;Wang S;Zha Z;Chen T;Deng T;Wang Y;Zhang T;Chen Y;Yuan Q;Zhao Q;Zhang J;Gu Y;Li S;Xia N
摘要当前的2019冠状病毒病（COVID-19）大流行是高致病性冠状病毒严重急性呼吸综合征冠状病毒2（SARS-CoV-2）快速传播的结果，对此没有有效的疫苗或治疗方法。为了开发疫苗，我们使用昆虫细胞表达系统表达并评估了刺突（S）蛋白的四种形式作为潜在候选物：受体结合结构域（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-2 P）。我们发现RBD在溶液中以单体形式出现，而S1、S-WT和S-2 P以具有大量糖基化的同源三聚体形式缔合。冷冻电子显微镜分析表明，S-2 P假设一个相同的三聚体构象的类似工程的S蛋白在293哺乳动物细胞中表达，但减少糖基化。总体而言，这四种蛋白质在酶联免疫吸附测定（ELISA）中赋予恢复期COVID-19患者血清优异的抗原性，但在免疫印迹中显示出不同的反应性。在三轮免疫方案后，RBD、S-WT和S-2 P而不是S1在小鼠中诱导高中和滴度（>3-log）。S-2 P的高免疫原性可在最低剂量（1 μg）下通过加入铝佐剂维持。 较高剂量（20 μg）的S-2 P可在非人灵长类动物中引起高中和滴度，超过在恢复期COVID-19受试者中测量的平均滴度的40倍。 我们的结果表明，来自昆虫细胞的融合前三聚体稳定的SARS-CoV-2 S蛋白可能为重组COVID-19疫苗的开发提供潜在的候选策略。
ABSTRACT 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.
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影响因子：
48
作者：
Kucukelbir, Alp;Sigworth, Fred J.;Tagare, Hemant D.
通讯作者：
Tagare, Hemant D.
DOI：
10.1038/nrmicro.2016.81
发表时间：
2016-08
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
de Wit E;van Doremalen N;Falzarano D;Munster VJ
通讯作者：
Munster VJ
影响因子：
13.2
作者：
Chan, Jasper Fuk-Woo;Kok, Kin-Hang;Yuen, Kwok-Yung
通讯作者：
Yuen, Kwok-Yung
DOI：
10.1007/978-1-0716-0373-4_16
发表时间：
2020-01-01
期刊：
EXPRESSION, PURIFICATION, AND STRUCTURAL BIOLOGY OF MEMBRANE PROTEINS
影响因子：
--
作者：
Bloch, Magnus;Santiveri, Nica;Taylor, Nicholas M. I.
通讯作者：
Taylor, Nicholas M. I.
DOI：
10.1073/pnas.1407087111
发表时间：
2014-10-21
影响因子：
11.1
作者：
Millet, Jean Kaoru;Whittaker, Gary R.
通讯作者：
Whittaker, Gary R.