Recombinant expression and immunogenicity evaluation of seasonal influenza virus hemagglutinin
10.3760/cma.j.cn112309-20240628-000235
- VernacularTitle:季节性流感病毒血凝素的重组表达与免疫原性评价
- Author:
Shenghua GUO
1
;
Yunpeng BAI
1
;
Yichi ZHANG
1
;
Xinming ZHANG
1
;
Changhao WANG
1
;
Chunping YAO
1
;
Yuanyuan LI
1
Author Information
1. 国药中生生物技术研究院（新型疫苗国家工程研究中心）疫苗研发课题组（一），北京 101111
- Publication Type:Journal Article
- Keywords:
Hemagglutinin;
Influenza virus;
Immunogenicity;
Insect-baculovirus expression system
- From:
Chinese Journal of Microbiology and Immunology
2025;45(8):680-686
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct four recombinant hemagglutinin (HA) antigens from seasonal influenza viruses and evaluate their immunogenicity in mouse models.Methods:HA coding sequences from four seasonal influenza virus strains Wisconsin (H1N1), Darwin (H3N2), Austria (B/Victoria lineage, BV) and Phuket (B/Yamagata lineage, BY) were optimized and synthesized, and then used to construct four recombinant plasmids. Recombinant baculoviruses were obtained through transformation and transfection. The expression of recombinant HA antigens was identified by SDS-PAGE and Western blot. The recombinant HA antigens were purified by nickel column affinity chromatography and intramuscularly administered to BALB/c mice after formulation with Al(OH) 3 or AddaVax adjuvant. Humoral immune responses were assessed by indirect ELISA and hemagglutination inhibition test, while cellular immune responses were evaluated by ELISPOT. Microneutralization test was used to detect the titers of serum antibodies in mice. Statistical analysis was performed using t test or non-parametric rank sum test. Results:PCR amplification and agarose gel electrophoresis confirmed the correct construction of the recombinant bacmids. Western blot showed verified the successful expression of the four recombinant antigens (H1-HA, H3N2-HA, BV-HA, and BY-HA). SDS-PAGE results showed that the purity of all four recombinant HA antigens exceeded 95%. After three-dose immunization, the total IgG levels in mice immunized with the recombinant H1N1-HA, H3N2-HA, or BV-HA formulated with AddaVax adjuvant were higher than those in the corresponding groups immunized with the same recombinant antigen alone (all P<0.05). The secretion levels of IFN-γ, IL-2, and IL-4 in the group receiving the mixture of all four recombinant HA antigens formulated with AddaVax adjuvant were higher than those in the group immunized with a commercial quadrivalent split influenza vaccine (all P<0.01). Results of the microneutralization test showed that the antibody titer in the quadrivalent split influenza vaccine group was 1∶225, whereas the titer in the group immunized with the mixture of four recombinant HA antigens formulated with AddaVax adjuvant could reach up to 1∶1 200. Conclusions:In this study, four recombinant seasonal influenza virus HA antigens are successfully expressed and demonstrated good immunogenicity in mice when formulated AddaVax adjuvant.
