Preliminary study on intranasal immunization of SARS-CoV-2 RBD protein delivered by Sendai virus to enhance respiratory mucosal and systemic immune response
Journal Title: Journal of Air Force Medical University - Year 2023, Vol 44, Issue 7
Abstract
Objective To study the characteristics of Sendai virus ( SeV) as a delivery vector in enhancing the respiratory mucosal and systemic immune response induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RBD protein. Methods The eukaryotic expression plasmid of SARS-CoV-2 RBD and antibody Fc fusion protein-coding gene was constructed by homologous recombination technology, and transfected into 293-T cells for expression and verified by Western blotting. The expression plasmid of RBD-Fc fusion protein was transfected into 293-T cells for large-scale expression and was purified by affinity chromatography. RBD-Fc protein was delivered by SeV for intranasal immunization, and RBD-Fc group, SeV group and PBS group were set as controls. Each group was given booster immunization once after 2 weeks, and the levels of specific antibodies in serum samples and bronchoalveolar lavage fluid (BALF) were detected after 3 weeks. Results The pCAGGS-RBD-Fc eukaryotic expression plasmid was successfully constructed by overlapping PCR to obtain the fusion protein gene fragment (RBD-Fc) of RBD and Fc protein-encoding gene fragments. Western blotting results showed that the expression band of RBD-Fc fusion protein was in line with the expected size. After large-scale expression, a single band close to Mr 60 000 was obtained by affinity chromatography, which was consistent with the expected size and high purity. Compared with RBD-Fc group, SeV group and PBS group, RBD-specific IgG could be detected in serum samples and BALF of mice immunized by SeV delivering RBD-Fc protein through nasal cavity ( serum: P < 0. 01, BALF: P < 0. 05), and RBD-specific IgA in BALF was higher than that in RBD-Fc group. Conclusion SeV can induce a certain level of mucosal and systemic immunity in delivering SARS-CoV-2 RBD protein for intranasal immunization, providing a preliminary experimental basis for the study of mucosal vaccines for respiratory infectious diseases.
Authors and Affiliations
YUAN Mingcheng, ZHANG Jian, CHEN Yang, XUE Pan, WU Fuxing, YUAN Ruodong, DONG Yangchao, XU Zhikai,ZHANG Fanglin, LEI Yingfeng
Keywords
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