[1]郭晓琴,张谞霄,马诚太,等.H7N9亚型禽流感病毒血凝素蛋白在杆状病毒中的表达及其免疫效力评估[J].江苏农业学报,2022,38(01):143-150.[doi:doi:10.3969/j.issn.1000-4440.2022.01.017]
　GUO Xiao-qin,ZHANG Xu-xiao,MA Cheng-tai,et al.Expression of hemagglutinin protein of H7N9 subtype avian influenza virus in baculovirus and evaluation of its immune efficacy[J].,2022,38(01):143-150.[doi:doi:10.3969/j.issn.1000-4440.2022.01.017]
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H7N9亚型禽流感病毒血凝素蛋白在杆状病毒中的表达及其免疫效力评估()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
38
期数:
2022年01期
页码:
143-150
栏目:
畜牧兽医·水产养殖
出版日期:
2022-02-28

文章信息/Info

Title:
Expression of hemagglutinin protein of H7N9 subtype avian influenza virus in baculovirus and evaluation of its immune efficacy
作者:
郭晓琴1; 2; 张谞霄2; 马诚太2; 欧杰1; 李柏林1; 李鑫2
（1.上海海洋大学食品学院，上海201306；2.勃林格殷格翰动物保健<中国>有限公司上海分公司，上海201203）
Author(s):
GUO Xiao-qin1; 2; ZHANG Xu-xiao2; MA Cheng-tai2; OU Jie1; LI Bai-lin1; LI Xin2
(1.College of Food Science, Shanghai Ocean University, Shanghai 201306, China;2.Boehringer Ingelheim Animal Health (China) Co., Ltd. Shanghai Branch, Shanghai 201203, China)
关键词:
H7N9亚型禽流感病毒; 重组杆状病毒; 血凝素; 免疫原性
Keywords:
H7N9 subtype avian influenza virus; recombinant baculovirus; hemagglutinin; immunogenicity
分类号:
S852.65+7
DOI:
doi:10.3969/j.issn.1000-4440.2022.01.017
文献标志码:
A
摘要:
旨在利用杆状病毒表达系统构建1株对高致病性H7N9亚型禽流感病毒（A/Chicken/Huizhou/HZ-3/2016）攻击后的家禽提供保护的候选疫苗株。用同源重组的方法构建1株表达H7N9亚型禽流感病毒（A/chicken/Shaoxing/5201/2013株）血凝素(Hemagglutinin, HA)蛋白的重组杆状病毒。用PCR技术鉴定重组杆状病毒rBac-SX5201HA的遗传稳定性，用间接免疫荧光方法和Western Blotting方法鉴定HA蛋白的表达情况，用血凝试验检测HA蛋白的体外活性，继而对重组疫苗在无特定病原体(Specific pathogen free, SPF)鸡上进行免疫效力试验，并对免疫后21 d的SPF鸡血清进行抗体检测，对攻毒5 d后的鸡咽喉和泄殖腔棉拭子进行病毒分离。结果显示，重组杆状病毒rBac-SX5201HA在昆虫细胞中生长良好，且可稳定高效表达H7 HA蛋白，血凝效价可达26；重组疫苗免疫SPF鸡21 d后可诱导28.4血凝抑制（Hemagglutinin inhibition, HI）抗体效价并能抵抗高致病性H7N9亚型禽流感病毒的攻击，免疫组SPF鸡群均未发病或死亡，仅有17%的SPF鸡在攻毒后第5 d出现排毒。可以看出，重组疫苗对高致病性H7N9亚型禽流感病毒攻击后的SPF鸡提供了100%的保护，且可有效抑制病毒在SPF鸡体内的复制。
Abstract:
The aim of this study is to construct a candidate vaccine strain for protection against highly pathogenic H7N9 subtype avian influenza virus （A/Chicken/Huizhou/HZ-3/2016） in poultry using baculovirus expression system. A recombinant baculovirus (rBac-SX5201HA) expressing H7N9 (A/chicken/Shaoxing/5201/2013) hemagglutinin (HA) protein was constructed. Genetic stability of rBac-SX5201HA was confirmed by PCR. Expression of HA protein was identified by indirect immunofluorescence assay (IFA) and Western blotting. The activity of HA protein in vitro was detected by HA assay. Immunogenicity and efficacy of recombinant vaccine were studied by animal trial on specific pathogen free (SPF) chickens. Antibody titer of chicken serum collected at 21 days after immunization was tested, and virus isolation was performed for chicken oropharyngeal and cloacal cotton swabs at five days post-challenge. The results showed that recombinant baculovirus (rBac-SX5201HA) stably and efficiently expressed H7 HA protein in insect cells, and HA titer could reach 26. The recombinant vaccine induced 28.4 hemagglutinin inhibition (HI) antibody titer in SPF chickens, and could resist the attack of highly pathogenic H7N9 subtype avian influenza virus. The SPF chickens in the immunized group did not get sick or die, and shedding was found from 17% SPF chickens at five days post-challenge. Recombinant vaccine provides protection against highly pathogenic H7N9 subtype avian influenza virus and effectively inhibits virus replication in SPF chickens.

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备注/Memo

备注/Memo:
收稿日期：2021-04-08基金项目：国家自然科学基金项目（31972188）作者简介：郭晓琴（1985-），女，江苏淮安人，硕士，研究方向为禽流感疫苗。(E-mail)guo1219@163.com通讯作者：李柏林,(E-mail)blli@shou.edu.cn；李鑫，(E-mail)xin_7.li@boehringer-ingelheim.com
更新日期/Last Update: 2022-03-04