[1]梁真洁?,潘俊慧?,于晓菲,等.H7N9禽流感DNA疫苗的免疫保护效力研究[J].中国预防兽医学报,2019,(09):935-939.[doi:0.3969/j.issn.1008-0589.201903018]
 LIANG Zhen-jie,PAN Jun-hui,YU Xiao-fei,et al.Protective efficacy of DNA vaccine against H7N9 avian influenza[J].Chinese journal of preventive veterinary medicine,2019,(09):935-939.[doi:0.3969/j.issn.1008-0589.201903018]
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H7N9禽流感DNA疫苗的免疫保护效力研究()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年09
页码:
935-939
栏目:
免疫学
出版日期:
2019-10-25

文章信息/Info

Title:
Protective efficacy of DNA vaccine against H7N9 avian influenza
文章编号:
1008-0589(2019)09-0935-05
作者:
 

梁真洁?潘俊慧?于晓菲陈普成曾显营柳金雄施建忠邓国华姜永萍*陈化兰*

 (中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室,哈尔滨 黑龙江 150069)
Author(s):
 

LIANG Zhen-jie? PAN Jun-hui? YU Xiao-fei CHEN Pu-cheng ZENG Xian-ying LIU Jin-xiong SHI Jian-zhong DENG Guo-hua JIANG Yong-ping* CHEN Hua-lan*

 (National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin 150069, China)
关键词:
H7N9禽流感DNA疫苗免疫保护效力
Keywords:
 H7N9 avian influenza  DNA vaccine  vaccination  protective efficacy
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201903018
文献标志码:
A
摘要:
为检测H7N9禽流感病毒(AIV) DNA疫苗的免疫保护效力,本研究将H7N9禽流感疫苗株A/Chicken/Guangxi/SD098/2017(H7N9) [CK/GX/SD098/17(H7N9)]的HA基因密码子优化后,定向克隆至载体pCAGGS中构建重组质粒pCA-SD098。将pCA-SD098转染至293T细胞后,采用间接免疫荧光(IFA)和western blot检测,结果显示,HA蛋白可以在293T细胞中正确表达。分别利用15 μg和20 μg重组质粒pCA-SD098免疫3周龄SPF鸡,3周后以相同的剂量和方式加强免疫,加强免疫一周后通过鼻腔分别感染105 EID50的同源高致病性AIV(HPAIV)CK/GX/SD098/17 (H7N9)和异源低致病性AIV(LPAIV) A/Chicken/Chongqing/SD057/2017(H7N9)[CK/CQ/SD057/17 (H7N9)],攻毒10 d内记录免疫鸡发病和死亡情况,检测免疫鸡HI抗体水平,并于攻毒后第3 d、第5 d和第7 d无菌采集所有鸡的喉头和泄殖腔拭子,采用红细胞凝集试验(HA)检测病毒的滴度。结果显示,重组质粒pCA-SD098免疫后可以诱导SPF鸡产生较高水平的HI抗体,15 μg pCA-SD098剂量免疫鸡后能够完全抵御H7N9 HPAIV 的致死性攻击,20 μg pCA-SD098剂量可以有效阻断H7N9 LPAIV的感染。攻毒后,所有免疫鸡无发病、无死亡、无排毒,本研究为质粒pCA-SD098作为防控H7N9禽流感的候选DNA疫苗提供了实验依据。
  
Abstract:
: In order to evaluate the immune protective efficacy of DNA vaccine against H7N9 avian influenza virus (AIV),We inserted the codon-optimized HA gene of the vaccine strain, A/Chicken/Guangxi/SD098/2017 (CK/GX/SD098/17, H7N9) into pCAGGs vector to generate a recombinant vector pCA-SD098. Indirect immunofluorescence assay and western blot analysis showed that the HA protein was correctly expressed in 293T cells transfected with the pCA-SD098 construct. Groups of 3-week-old SPF chickens were inoculated intramuscularly with 15μg or 20μg of pCA-SD098, and were boosted with the same dose and manner three weeks after the primary immunization. One week post-booster, the immunized chickens were challenged intranasally with 105 EID50 of homologous highly pathogenic avian influenza virus (HPAIV) CK/GX/SD098/17(H7N9) or heterologous low pathogenic avian influenza virus (LPAIV) A/Chicken/Chongqing/SD057/2017 (CK/CQ/SD057/17, H7N9). The signs of disease and death of experimental chickens were recorded for ten days, and the level of the HI antibodies was detected at different time post-challenge. Oropharyngeal and cloacal swabs were collected on day 3, 5, and 7 post-challenge for virus titration by hemagglutination assay (HA). The results showed that recombinant plasmid pCA-SD098 induced high level of HI antibodies, 15 μg pCA-SD098 protected chickens from lethal challenge by the H7N9 HPAIV, and 20 μg pCA-SD098 completely blocked the infection by the H7N9 LPAIV. No signs of disease, death, or virus shedding were observed in the immunized chickens. These data indicate that the recombinant plasmid pCA-SD098 has the potential to be a DNA vaccine candidate against H7N9 AIV. Our study provides an experimental basis for the prevention and control of H7N9 avian influenza.

参考文献/References:

[1] Lin Ting-hui, Chia M Y, Lin Chun-yang, et al. Improving immunogenicity of influenza virus H7N9 recombinant hemagglutinin for vaccine development [J]. Vaccine, 2019, 37(13): 1897-1903.
[2]Shi Jian-zhong, Deng Guo-hua, Kong Hui-hui, et al. H7N9 virulent mutants detected in chickens in China pose an increased threat to humans [J]. Cell Res, 2017, 27(12): 1409-1421.
[3]Shi Jian-zhong, Deng Guo-hua, Ma Shu-jie, et al. Rapid evolution of H7N9 highly pathogenic viruses that emerged in China in 2017 [J]. Cell Host & Microbe, 2018, 24(4): 558-568.
[4]王祥,周东明. 基于血凝素(HA)的新型流感疫苗研究进展[J]. 生命科学,2014,26(09):943-948.
[5]舒黛廉,王慧杰,任敏. 核酸疫苗研究现状[J]. 中国畜牧兽医,2005,(10):43-45.
[6]Yang Lei, Zhu Wen-fei, Li Xi-yan, et al. Genesis and spread of newly emerged highly pathogenic H7N9 avian viruses in MAinland China [J]. J Virol, 2017, 91(23): 1-13.
[7]Chen Yu, Liang Wei-feng, Yang Shi-gui, et al. Human infections with the emerging avian influenza A H7N9 virus from wet market poultry: clinical analysis and characterisation of viral genome [J]. Lancet, 2013, 381(9881): 1916-1925.
[8]Zeng Xian-ying, Tian Guo-bin, Shi Jian-zhong, et al. Vaccination of poultry successfully eliminated human infection with H7N9 virus in China [J]. Sci China Life Sci, 2018, 61(12): 1465-1473.
[9]Imai M, Watanabe T, Kiso M, et al. A Highly Pathogenic avian H7N9 influenza virus isolated from a human is lethal in some ferrets infected via respiratory droplets [J]. Cell Host Microbe, 2017, 22(5): 615-626.
[10]黄冬,陈普成,刘兵,等. 用于构建双价禽流感DNA疫苗的双目的基因表达质粒的构建及鉴定[J]. 中国预防兽医学报,2016,38(04):257-261.
[11]王国俊,熊杰,李俊平,等. H5亚型禽流感病毒A/Duck/ Anhui/1/2006(Clade 2.3)密码子优化HA基因DNA疫苗的免疫保护效力研究[J]. 中国预防兽医学报,2010,32(02):116-120.
[12]Jiang Yong-ping, Yu Kang-zhen, Zhang Hong-bo, et al. Enhanced protective efficacy of H5 subtype avian influenza DNA vaccine with codon optimized HA gene in a pCAGGS plasmid vector [J]. Antiviral Res, 2007, 75(3): 234-241.
[13]Ltd V P, Park B, Adelaide, et al. Molecular mechanisms for enhanced DNA vaccine immunogenicity [J]. Expert Rev Vaccines, 2016, 15(3): 313-329.
(本文编辑:李 娜;英文编辑:李呈军)
 

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更新日期/Last Update: 2019-10-29