新孢子虫与弓形虫交叉抗原AMA1基因重组腺病毒构建及免疫应答分析
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- 论文作者：贾立军 ; 李航 ; 张贺洋 ; 姜利建 ; 刘晋宇 ; 兰岚 ; 凌芳芳 ; 石甜甜 ; 赵昕月
- 年：2016
- 作者机构：延边大学农学院动物医学系;
- 论文关键词：新孢子虫 ; 弓形虫 ; AMA1基因 ; 重组腺病毒 ; 免疫应答
- 会议召开时间：2016-07-20
- 会议录名称：中国畜牧兽医学会兽医公共卫生学分会第五次学术研讨会论文集
- 语种：中文
- 分类号：S852.7
- 学会代码：SYGW
- 会议名称：中国畜牧兽医学会兽医公共卫生学分会第五次学术研讨会
- 会议地点：中国内蒙古通辽
- 主办单位：中国畜牧兽医学会兽医公共卫生学分会
- 学会名称：中国畜牧兽医学会兽医公共卫生学分会
- 页数：5
- 文件大小：644k
- 原文格式：D
- 会议级别：全国
摘要
为构建新孢子虫和弓形虫AMA1基因重组腺病毒,并分析其免疫原性,本试验根据新孢子虫和弓形虫AMA1基因序列的开放阅读框,设计新孢子虫和弓形虫交叉抗原AMA1基因通用引物,构建重组克隆质粒p MD18T-Nc AMA1和p MD18T-Tg AMA1、重组腺病毒穿梭质粒ADV4-Nc/Tg AMA1,将ADV4-Nc/Tg AMA1和骨架质粒pac Ad5线性化后共转染293T细胞,包装Ad5-Nc/Tg AMA1重组腺病毒,测定病毒滴度后,收集病毒液接种BALB/c小鼠,间接ELISA检测小鼠血清Ig G抗体水平。结果显示,Nc/Tg AMA1基因在Ad5-Nc/Tg AMA1重组腺病毒中获得表达,测定Ad5-Nc/Tg AMA1重组腺病毒滴度为109 PFU/m L,接种BALB/c小鼠后,Ad5-Nc/Tg AMA1接种组Ig G抗体水平极显著高于p VAX1-Nc/Tg AMA1质粒组(P<0.01)和PBS对照组(P<0.01)。说明构建的Ad5-Nc/Tg AMA1重组腺病毒能够诱导小鼠产生特异性体液免疫应答,本试验为新孢子虫和弓形虫交叉抗原AMA1基因重组腺病毒载体疫苗的研制奠定了基础。
引文
[1]Zhang G,Huang X,Boldbaatar D,et al.Cons truction of Neospora caninum stably expressing TgSAG1 and evaluation of its protective effects against Toxoplasma gondii infection in mice[J].Vaccine,2010,28(45):7243-7247.
[2]孙婷婷,金京顺,贾立军.新孢子虫与弓形虫交叉抗原基因AMA1的克隆及原核表达载体构建[J].延边大学农学学报,2015,37(1):8-11.
[3]Yu L,Yamagishi J,Zhang S,et al.Protective effect of a prime-boost strategy with plasmid DNA followed by recombinant adenovirus expressing Tg AMA1 as vaccines against Toxoplasma gondii infection in mice[J].Parasit Int,2012,61(3):481-486.
[4]Zhang H,Nishikawa Y,Yamagishi J,et al.Neospora caninum:application of apical membrane antigen 1encapsulated in the oligomannose-coated liposomes for reduction of offspring mortality from infection in BALB/c mice[J].Exp Parasitol,2010,125(2):130-136.
[5]Zhang H,Compaore M K,Lee E G,et al.Apical membrane antigen 1 is a cross-reactive antigen between Neospora caninum and Toxoplasma gondii,and the anti-Nc AMA1 antibody inhibits host cell invasion by both parasites[J].Mol Biochem Parasit,2007,151(2):205-212.
[6]Morishita R,Aoki M,Hashiya N,et al.Safety evaluation of clinical gene therapy using hepatocyte growth factor to treat peripheral arterial disease[J].Hypertension,2004,44(2):203-209.
[7]郭焕平.牛源犬新孢子虫AMA1基因核酸疫苗与重组腺病毒疫苗的制备及动物免疫[D].延吉:延边大学,2014.
[8]潘俊杰,施海明,罗心平,等.U937单核细胞几种不同转染方法的比较[J].复旦学报(医学版).2010,37(5):594-596.
[9]Wraith D C,Vessey A E,Askonas B A.Purified influenza virus nucleoprotein protects mice from lethal infection[J].J Gen Virol,1987,68(2):433-440.
[10]贾立军,张守发,鲁承.犬新孢子虫可溶性抗原分析及间接ELISA检测方法的建立[J].中国兽医杂志,2009,45(9):25-26.
[11]陈千林,王振宝,吉尔格力,等.基于Nc2和Nc5基因的新孢子虫PCR方法的建立[J].中国畜牧兽医,2016,43(3):622-628.
[12]Sayles P C,Gibson G W,Johnson L L.B cells are essential for vaccination induced resistance to virulent Toxoplasma gondii[J].Infect Immun,2000,68(3):1026-1033
[13]Wurtz O,Bajenoff M,Guerder S.IL-4-mediated inhibition of IFN-γproductionby CD4+T cells proceeds by several developmentally regulated mechanisms[J]Int Immunol,2004,16(3):501-508.
[14]Lindsay DS,Lenz SD,Blagburn BL,et al.Characterization of mperature-sn sitive strains of Neospora caninum in mice[J]J Parasitol,1999,85(1):64-67.
[15]Walsh CP,Vemulapalli R,Sriranganathan N,et al.Molecular comparison of the dense granule proteins GRA6and GRA7 of Neospora hughesi and Neospora caninum[J].Int J Parasitol,2001,31(3):253-258.
[16]王素华.腺病毒载体及其在基因治疗中的应用[J].中国畜牧兽医,2009,36(9):119-121.
[2]孙婷婷,金京顺,贾立军.新孢子虫与弓形虫交叉抗原基因AMA1的克隆及原核表达载体构建[J].延边大学农学学报,2015,37(1):8-11.
[3]Yu L,Yamagishi J,Zhang S,et al.Protective effect of a prime-boost strategy with plasmid DNA followed by recombinant adenovirus expressing Tg AMA1 as vaccines against Toxoplasma gondii infection in mice[J].Parasit Int,2012,61(3):481-486.
[4]Zhang H,Nishikawa Y,Yamagishi J,et al.Neospora caninum:application of apical membrane antigen 1encapsulated in the oligomannose-coated liposomes for reduction of offspring mortality from infection in BALB/c mice[J].Exp Parasitol,2010,125(2):130-136.
[5]Zhang H,Compaore M K,Lee E G,et al.Apical membrane antigen 1 is a cross-reactive antigen between Neospora caninum and Toxoplasma gondii,and the anti-Nc AMA1 antibody inhibits host cell invasion by both parasites[J].Mol Biochem Parasit,2007,151(2):205-212.
[6]Morishita R,Aoki M,Hashiya N,et al.Safety evaluation of clinical gene therapy using hepatocyte growth factor to treat peripheral arterial disease[J].Hypertension,2004,44(2):203-209.
[7]郭焕平.牛源犬新孢子虫AMA1基因核酸疫苗与重组腺病毒疫苗的制备及动物免疫[D].延吉:延边大学,2014.
[8]潘俊杰,施海明,罗心平,等.U937单核细胞几种不同转染方法的比较[J].复旦学报(医学版).2010,37(5):594-596.
[9]Wraith D C,Vessey A E,Askonas B A.Purified influenza virus nucleoprotein protects mice from lethal infection[J].J Gen Virol,1987,68(2):433-440.
[10]贾立军,张守发,鲁承.犬新孢子虫可溶性抗原分析及间接ELISA检测方法的建立[J].中国兽医杂志,2009,45(9):25-26.
[11]陈千林,王振宝,吉尔格力,等.基于Nc2和Nc5基因的新孢子虫PCR方法的建立[J].中国畜牧兽医,2016,43(3):622-628.
[12]Sayles P C,Gibson G W,Johnson L L.B cells are essential for vaccination induced resistance to virulent Toxoplasma gondii[J].Infect Immun,2000,68(3):1026-1033
[13]Wurtz O,Bajenoff M,Guerder S.IL-4-mediated inhibition of IFN-γproductionby CD4+T cells proceeds by several developmentally regulated mechanisms[J]Int Immunol,2004,16(3):501-508.
[14]Lindsay DS,Lenz SD,Blagburn BL,et al.Characterization of mperature-sn sitive strains of Neospora caninum in mice[J]J Parasitol,1999,85(1):64-67.
[15]Walsh CP,Vemulapalli R,Sriranganathan N,et al.Molecular comparison of the dense granule proteins GRA6and GRA7 of Neospora hughesi and Neospora caninum[J].Int J Parasitol,2001,31(3):253-258.
[16]王素华.腺病毒载体及其在基因治疗中的应用[J].中国畜牧兽医,2009,36(9):119-121.
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