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

doi:10.13523/j.cb.2203003

China Biotechnology

2022, Vol. 42

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

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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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

Received: 02 March 2022 Published: 17 June 2022

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Corresponding Authors: Ya-feng LI E-mail: muran2001@163.com

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	Man-yun QIAN
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Cite this article:

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.

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

Fig.1 Plasmid map of SARS-CoV-2 S1 and S (a) Plasmid (pcDNA3.1-nCoV-S1B) map of SARS-CoV-2 S1 (b) Plasmid (pcDNA3.1-nCoV-SF4) map of SARS-CoV-2 S

Fig.2 A picture of the sodium dodecyl sulfate polyacrylamide gel electrophoresis from SARS-CoV-2 S1 and S protein (a) A picture of the sodium dodecyl sulfate polyacrylamide gel electrophoresis from SARS-CoV-2 S1 protein (b) A picture of the sodium dodecyl sulfate polyacrylamide gel electrophoresis from SARS-CoV-2 S protein

Table 1 Recombinant SARS-CoV-2 S1 and S protein vaccines

Fig.3 Serum IgG antibody titers in the immunized mice detected by ELISA (a) The absorbances of sera of the immunized mice with S1 or S protein (S1 dilution 1∶150, S dilution 1∶450) measured at 450 - 630 nm (b) Serum IgG titers of mice inoculated with S1 or S protein (n = 5. ^**, P<0.01; ^***, P<0.001; ^****, P<0.000 1)

Fig. 4 Effects of different doses of SARS-CoV-2 S1 and S proteins on neutralizing antibody production in immunized mice (a-f) Respectively represent the neutralizing activity titers of mouse serum immunized with different doses of S1 and S protein against SARS-CoV-2-Fluc WT, B.1.1.7, P.1, B.1.617.2, B.1.621, 501Y.V2-1 strains (n=5. ^*, P<0.05; ^**, P<0.01; ^***, P<0.001; ^****, P<0.000 1)

Fig.5 Serum neutralizing titers in the immunized mice against different pseudotype SARS-CoV-2 strains Serum neutralizing titers of the mice immunized with 5 μg of S1 protein and 10 μg of S protein against SARS-CoV-2-Fluc WT, B.1.1.7, P.1, B.1.617.2, B.1.621 and 501Y.V2-1 strains (n=5; ^*, P<0.05, ^**, P< 0.01, ^***, P< 0.001; ^****, P< 0.000 1)

Fig.6 Cellular immune response in the spleen cells of the immunized mice induced by the recombinant SARS-CoV-2 S1 and S proteins vaccine candidates (a),(b) The levels of IFN-γ, IL-4 induced by mouse specific spleen cells detected by ELISpot (n=5; ^*, P<0.05; ^**, P<0.01; ^***, P< 0.001; ^****, P< 0.000 1)

Table S1 Amino acid sequences of S1 and S proteins of SARS-CoV-2


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