Prokaryotic expression of bovine coronavirus N protein and preliminary evaluation of its immunological effect on mice
10.16303/j.cnki.1005-4545.2024.12.06
- VernacularTitle:牛冠状病毒N蛋白的原核表达及对小鼠免疫原性的初步评价
- Author:
Qian JIANG
1
;
Fang MIN
;
Zhigang MA
;
Yumeng LIANG
;
Xinyu TAO
;
Xiaojun DING
;
Tian-qing LI
;
Qi ZHONG
;
Gang YAO
;
Xuelian MA
Author Information
1. 新疆农业大学动物医学学院新疆草食动物新药研究与创制重点实验室,新疆乌鲁木齐 830052
- Publication Type:Journal Article
- Keywords:
BCoV;
N protein;
subunit vaccine;
mice;
immunogenicity
- From:
Chinese Journal of Veterinary Science
2024;44(12):2540-2548
- CountryChina
- Language:Chinese
-
Abstract:
Bovine coronavirus(BCoV)can cause diarrhea in calves,as well as respiratory diseases,and lead to the death of calves in severe cases,resulting in great economic losses to the cattle pro-duction.BCoV is highly prevalent and widely spread.So far in China there is no independently de-veloped vaccine against BCoV on the market.In this study,the most conserved N gene of BCoV was first selected to construct a prokaryotic expression vector,and the N protein was expressed and purified;subsequently,BALB/c mice were immunized with BCoV N protein combined with Freund's adjuvant,and the antibody potency,the proportion of splenic T-lymphocyte subpopulation and the cytokine release of the immunized mice were detected by ELISA and flow cytometry.The results showed that a soluble BCoV N protein about 55 kDa in size was successfully obtained.The ELISA test showed that the antibody potency of serum IgG and IgA was 1∶51 200 and 1∶3 200,respectively,in mice immunized with BCoV N protein combined with Freund's adjuvant.The flow cytometry test showed that the percentage of CD4+/CD8+T-lymphocytc subsets was extremely higher in the immunized mice(P＜0.01),and the release of TNF-α was significantly higher(P＜0.05),producing a cellular immune response biased towards the Th1 type in comparison with that in control mice.It is indicated that the soluble expression of BCoV N protein can be successfully a-chieved by the prokaryotic expression system,and the obtained BCoV N protein presents good im-munogenicity which induces strong humoral,mucosal and cellular immune responses in the immu-nized mice.Our study provides an important technical support for the development of a safe and ef-fective BCoV subunit vaccine.
