Preparation and immunogenicity evaluation of recombinant influenza hemagglutinin trimer vaccine
10.3760/cma.j.cn112309-20220926-00312
- VernacularTitle:重组流感血凝素三聚体蛋白疫苗的制备及免疫原性评价
- Author:
Guomei ZHANG
1
;
Jing LIU
;
Ning MA
;
Rong ZHOU
;
Yang LE
;
Zhegang ZHANG
;
Xuanxuan NIAN
;
Xuedan LI
;
Jiayou ZHANG
;
Yangyang ZHANG
;
Sheng LI
;
Hu HUANG
;
Xiaoming YANG
Author Information
1. 武汉生物制品研究所有限责任公司病毒性疫苗研究二室，武汉 430207
- Keywords:
Influenza virus;
Genetically engineered vaccine;
Recombinant subunit vaccine;
Hemagglutinin;
Trimeric protein
- From:
Chinese Journal of Microbiology and Immunology
2023;43(2):137-143
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare a recombinant hemagglutinin trimer (HA-Tri) vaccine against influenza viruses and to study its immunogenicity in a mouse model.Methods:A stable CHO cell line that could express HA-Tri was constructed. Western blot, single radial immunodiffusion, protein particle size detection and N-glycosylation site analysis were performed for qualitative and quantitative analysis of the recombinant protein. According to the different treatment conditions such as dosage and adjuvant, BALB/c mice were divided into 11 groups and subjected to consistent immunization procedures. Serum neutralizing antibody titers were measured on 56 d after the first immunization to evaluate the immunogenicity of HA-Tri.Results:The constructed CHO cells could secret and express HA-Tri proteins. The HA-Tri proteins were biologically active and capable of forming precipitation rings in the single radial immunodiffusion. The particle size of HA-Tri was approximately 18.79 nm and 10 N-glycosylation sites were detected, including high mannose, complex glycoforms and heterozygous glycoforms. After prime-boost immunization, there was no statistically significant difference in the titers of neutralizing antibodies induced in mice by 3.75 μg of HA-Tri in combination with RFH01 adjuvant and 15 μg of monovalent vaccine stock solution ( P=0.431 2, U=36). Serum antibody titers in the HA-Tri+ RFH01 groups were higher than those in the corresponding HA-Tri groups without RFH01 adjuvant, and the highest titer was induced in the 15 μg HA-Tri+ RFH01 group, which was 1 280. Conclusions:The recombinant HA-Tri protein was successfully prepared. HA-Tri in combination with RFH01 adjuvant could induce humoral immune responses against influenza viruses in BALB/c mice, which would provide reference for the development of influenza virus recombinant subunit vaccines.
