[1]王晓雪,刘延珂,杨东东,等.表达PEDV中国变异株S蛋白复制型重组人腺病毒4型的构建及其免疫原性研究[J].中国预防兽医学报,2018,(02):127-131.[doi:0.3969/j.issn.1008-0589.201704040]
 WANG Xiao-xue,LIU Yan-ke,YANG Dong-dong,et al.Construction and immunogenicity of replicating recombinant human adenovirus serotype 4 expressing S protein of PEDV Chinese variant strain[J].Chinese journal of preventive veterinary medicine,2018,(02):127-131.[doi:0.3969/j.issn.1008-0589.201704040]
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表达PEDV中国变异株S蛋白复制型重组人腺病毒4型的构建及其免疫原性研究()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年02
页码:
127-131
栏目:
免疫学
出版日期:
2018-02-15

文章信息/Info

Title:
Construction and immunogenicity of replicating recombinant human adenovirus serotype 4 expressing S protein of PEDV Chinese variant strain
文章编号:
1008-0589(2018)2-0127-06
作者:
 

王晓雪刘延珂杨东东吴艳阳高冬生王永生李永涛赵 军*王川庆张改平

 (河南农业大学 牧医工程学院,河南 郑州 450002)
Author(s):
 

WANG Xiao-xue LIU Yan-ke YANG Dong-dong WU Yan-yang GAO Dong-sheng WANG Yong-sheng LI Yong-tao ZHAO Jun* WANG Chuan-qing ZHANG Gai-ping

 (College of Animal Science and Veterinary Science, Henan Agricultural University, Zhengzhou 450002, China)
关键词:
 猪流行性腹泻病毒S基因复制型重组人腺病毒血清4型活载体疫苗
Keywords:
 Porcine epidemic diarrhea virus  S gene  replicativerecombinant human adenovirus serotype 4  live vector vaccine
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201704040
文献标志码:
A
摘要:
为构建表达猪流行性腹泻病毒(PEDV) S蛋白复制型重组人腺病毒4型(HAdV-4)及评价其免疫原性,本研究利用RT-PCR扩增得到PEDV中国变异株纤突(S)基因的N-末端区域(1 bp~1 140 bp),构建重组质粒pAd4FAST-Shuttle-S,重组质粒经线性化和同源重组,得到含有PEDV S基因的重组HAdV-4感染性克隆,纯化的感染性克隆DNA经PacⅠ线性化后转染HEK293细胞,拯救得到表达PEDV变异株S蛋白的重组病毒rAd4△E3-S。将该重组病毒免疫小鼠,间接ELISA测定其血清中抗S蛋白抗体水平,利用体外淋巴细胞转化试验和流式细胞技术检测免疫小鼠T淋巴细胞反应。结果显示,重组病毒能够表达PEDV S蛋白,并且能刺激小鼠产生特异性体液和细胞免疫应答。本实验结果为进一步研究重组病毒在猪体上的免疫反应奠定基础,也为PEDV活载体疫苗的研发提供实验依据。
Abstract:
  In order to construct a replicative recombinant human adenovirus serotype 4 (HAdV-4) expressing the S protein of PEDV and evaluate the immunogenicity of the recombinant virus. The N-terminal of S gene (1 bp-1 140 bp) in PEDV Chinese variant strain was cloned into shuttle vector pAd4FAST-Shuttle. The constructive recombinant plasmid pAd4FAST-Shuttle-S was linearized, and transformed into BJ5183 host harboring HAdV-4 backbone plasmid for homologous recombination to get the infectious clone of HAdV-4 containing the PEDV S gene. The linearized infectious clone DNA was transfected into HEK293 cells to rescue the recombinant virus rAd4△E3-S, and the immunogenicity of rAd4△E3-S was evaluated with mouse model. Antibody responses against PEDV S protein were determined by indirect ELISA, and the T lymphocyte responses were evaluated by lymphocyte proliferation assay and flow cytometry. The results showed that the rAd4△E3-S could induce specific humoral and cellular immune responses against PEDV spntein and the results might lay a foundation for clinical evaluationin of the Ad4△E3-S with swine model.

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(本文编辑:李 爽)

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更新日期/Last Update: 2018-03-09