SARS-CoV-2重组S1和S蛋白疫苗诱导保护性免疫的研究<sup>*</sup>

doi:10.13523/j.cb.2203003

中国生物工程杂志

2022, Vol. 42

Issue (5): 106-116 DOI: 10.13523/j.cb.2203003

新冠肺炎疫苗的研究策略

SARS-CoV-2重组S1和S蛋白疫苗诱导保护性免疫的研究^*

钱曼云¹,王继伟²,李颢泽²,王瑞华²,刘云²,李亚峰^3,^4,^5,^**(

)

1 山西医科大学基础医学院生物化学与分子生物学教研室太原 030001
2 南京诺唯赞生物科技股份有限公司南京 210000
3 山西医科大学第五临床医学院(山西省人民医院)中心实验室太原 030012
4 肾脏病山西省重点实验室太原 030012
5 山西医科大学微生态研究院太原 030001

Study on Protective Immunity Induced by Recombinant SARS-CoV-2 S1 and S Protein Vaccine

QIAN Man-yun¹,WANG Ji-wei²,LI Hao-ze²,WANG Rui-hua²,LIU Yun²,LI Ya-feng^3,^4,^5,^**(

)

1 Department of Biochemistry and Molecular Biology,College of Basic Medicine, Shanxi Medical University, Taiyuan 030001, China
2 Nanjing Vazyme Biotechnology Co.Ltd., Nanjing 210000, China
3 The Fifth Hospital of Shanxi Medical University (Shanxi Provincial People’s Hospital), Taiyuan 030012, China
4 Shanxi Key Laboratory of Nephrology, Taiyuan 030012, China
5 Institute of Microecology, Shanxi Medical University, Taiyuan 030001, China

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摘要：

目的:评价新型冠状病毒(SARS-CoV-2)重组S1蛋白和S蛋白疫苗对SARS-CoV-2的免疫保护效果。方法:将SARS-CoV-2重组S1蛋白和S蛋白分别联合氢氧化铝佐剂以0.1 μg/只、1 μg/只、5 μg/只、10 μg/只不同剂量接种6~8周BALB/c纯系健康雌性小鼠。第二次免疫后采血通过酶联免疫吸附试验(ELISA)检测血清中IgG抗体效价,通过假病毒中和试验比较免疫小鼠血清对SARS-CoV-2野生型株(WT)、英国株(B.1.1.7)、巴西株(P.1)、印度株(B.1.617.2)、Mu毒株(B.1.621)和南非株(501Y.V2-1)六种假病毒毒株中和活性效价,取脾细胞通过酶联免疫斑点技术(ELISpot)检测免疫小鼠的细胞免疫水平。结果:SARS-CoV-2重组S和S1蛋白都能诱导小鼠产生较强的IgG抗体水平。免疫S1蛋白的小鼠血清对SARS-CoV-2野生型株、英国株、巴西株有明显的中和活性,免疫S蛋白的小鼠血清除了对SARS-CoV-2野生型株、英国株、巴西株有明显中和活性之外,对印度株也有明显的中和活性,两种蛋白质免疫的小鼠血清均对野生型株中和效果最强。S蛋白免疫的小鼠脾细胞能够显著诱导出γ干扰素(IFN-γ)和白介素-4(IL-4)的产生。S蛋白诱导产生的IgG抗体、中和抗体、细胞免疫水平均高于S1。结论:SARS-CoV-2重组S蛋白疫苗能够诱导产生较强的保护性免疫应答。

关键词： 新型冠状病毒; S蛋白; S1蛋白; 新型冠状病毒突变株; 保护性免疫应答

Abstract:

To evaluate the immune protection of recombinant SARS-CoV-2 S1 and S protein vaccine. Methods: Recombinant SARS-CoV-2 S1 or S protein combined with aluminum hydroxide adjuvant was inoculated at different doses of 0.1 μg, 1 μg, 5 μg and 10 μg per mouse for 6-8 weeks. Serum IgG antibody titers were detected by enzyme linked immunosorbent assay (ELISA) after second immunization. The serum neutralizing antibody titers of the immunized mice against pseudotype SARS-CoV-2-Fluc WT, B.1.1.7, P.1, B.1.617.2, B.1.621, 501Y.V2-1 strains were compared by pseudovirus neutralization test. The cellular immune levels of sera were detected by enzyme-linked immunospot assay (ELISpot).Results: Both SARS-CoV-2 S and S1 proteins could induce strong IgG antibody levels in mouse model. The sera of mice immunized with S1 protein showed obvious neutralization activity against SARS-CoV-2-Fluc WT, B.1.1.7 and P.1. The sera of mice immunized with the recombinant S protein also showed obvious neutralization activity against SARS-CoV-2-Fluc B.1.617.2 in addition to SARS-CoV-2-Fluc WT, B.1.1.7 and P.1. The serum of mice immunized with two kinds of proteins had the strongest neutralizing effect on SARS-CoV-2-Fluc WT. Mouse spleen cells immunized with S protein could significantly induce the production of interferon-γ (IFN-γ) and interleukin-4 (IL-4). The levels of IgG antibody, neutralizing antibody and cellular immunity induced by S protein were higher than those of S1.Conclusion: Recombinant SARS-CoV-2 S protein vaccine can induce protective immune responses.

Key words: SARS-CoV-2 S protein S1 protein SARS-CoV-2 mutant Protective immune responses

收稿日期: 2022-03-02 出版日期: 2022-06-17

ZTFLH:

Q819

基金资助: *山西省卫生健康委科研课题计划任务书(15)资助项目

通讯作者: 李亚峰 E-mail: muran2001@163.com

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引用本文:

钱曼云,王继伟,李颢泽,王瑞华,刘云,李亚峰. SARS-CoV-2重组S1和S蛋白疫苗诱导保护性免疫的研究^*[J]. 中国生物工程杂志, 2022, 42(5): 106-116.

QIAN Man-yun,WANG Ji-wei,LI Hao-ze,WANG Rui-hua,LIU Yun,LI Ya-feng. Study on Protective Immunity Induced by Recombinant SARS-CoV-2 S1 and S Protein Vaccine. China Biotechnology, 2022, 42(5): 106-116.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2203003 或 https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I5/106

图1 SARS-CoV-2 S1和S质粒图

图2 SARS-CoV-2 S1蛋白和S蛋白SDS-PAGE结果图

表1 SARS-CoV-2重组S1蛋白和S蛋白疫苗

图3 酶联免疫吸附试验检测小鼠血清中IgG抗体效价

图4 不同剂量的SARS-CoV-2重组S1蛋白和S蛋白对免疫小鼠中和抗体产生的影响

图5 免疫小鼠血清对不同SARS-CoV-2株的中和活性效价

图6 SARS-CoV-2重组S1蛋白和S蛋白免疫小鼠脾细胞产生的细胞免疫反应

表S1 SARS-CoV-2 S1蛋白和S蛋白氨基酸序列

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