Immune effect of H9N2 subtype AIV NP protein by prokaryotic expression
10.16303/j.cnki.1005-4545.2024.06.04
- VernacularTitle:H9N2亚型AIV NP蛋白的原核表达及其免疫效果
- Author:
Xiaofeng LI
1
;
Zhixun XIE
;
Zhihua RUAN
;
Meng LI
;
Dan LI
;
Minxiu ZHANG
;
Zhiqin XIE
;
Sisi LUO
;
You WEI
;
Liji XIE
;
Tingting ZENG
;
Yanfang ZHANG
;
Jiaoling HUANG
;
Sheng WANG
Author Information
1. 广西壮族自治区兽医研究所 农业农村部中国(广西)-东盟跨境动物疫病防控重点实验室/广西兽医生物技术重点实验室,广西南宁 530001
- Keywords:
H9N2 subtype avian influenza;
NP protein;
prokaryotic expression;
cytokines
- From:
Chinese Journal of Veterinary Science
2024;44(6):1113-1119
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P＜0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P＞0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.
