Construction and analysis of the immune response of Neospora caninum and Toxoplasma gondii cross recombinant adenovirus antigen AMA1 vaccine
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- 英文论文题名：Construction and analysis of the immune response of Neospora caninum and Toxoplasma gondii cross recombinant adenovirus antigen AMA1 vaccine
- 论文作者：贾立军 ; Shoufa ZHANG ; Longzheng YU ; Shujiang XUE ; Peng ZHAO
- 英文论文作者：Lijun JIA ; Shoufa ZHANG ; Longzheng YU ; Shujiang XUE ; Peng ZHAO ; Department of Veterinary Medicine ; Agriculture college of Yanbian University
- 年：2016
- 作者机构：Department of Veterinary Medicine, Agriculture college of Yanbian University;
- 英文论文关键词：Neospora caninum ; Toxoplasma gondii ; AMA1 gene ; recombinant adenovirus ; immune response
- 会议召开时间：2016-08-08
- 会议录名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会论文摘要集
- 英文会议录名称：The Abstracts of the Tenth Symposium for Yong Scientists of China Society of Parasitology
- 语种：英文
- 分类号：S852.723
- 学会代码：JISC
- 会议名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会
- 会议地点：中国四川成都
- 主办单位：中国动物学会寄生虫学专业青年委员会
- 学会名称：中国动物学会寄生虫学专业委员会
- 页数：1
- 文件大小：338k
- 原文格式：D
- 会议级别：全国
摘要
In order to establish AMA1 recombinant adenovirus of Neospora caninum and Toxoplasma gondii, and analysis the immunogenicity of it. According to the open reading frame of N. caninum and T. gondii AMA1 gene sequences, cross universal primers were designed. Based on p MD18T-Nc AMA1 and p MD18T-Tg AMA1 cloning plasmid, recombinant adenovirus shuttle plasmid ADV4-Nc/Tg AMA1 was constructed. Then, ADV4-Nc/Tg AMA1 and pac Ad5 backbone plasmid were linearized and co-transfected 293 T cells. After packaging recombinant adenovirus and measuring the virus titer, collected virus was inoculated into BALB/c mice, confirmed the Ig G antibody levels by indirect ELISA method. The results showed, Nc/Tg AMA1 gene was expressed in Ad5-Nc/Tg AMA1 recombinant adenovirus, Ad5-Nc/Tg AMA1 recombinant adenovirus titer was 109 PFU / mL. Ig G antibody levels in the Ad5-Nc/Tg AMA1 vaccinated inoculating group was significantly higher than p VAX1-Nc/Tg AMA1 plasmid group(P<0.01) and PBS control group(P<0.01). This result indicated that the constructed Ad5-Nc/Tg AMA1 recombinant adenovirus could induce specific humoral immune response in mice, this research laid a solid foundation for the development of a recombinant adenovirus vaccine against N. caninum and T. gondii.
In order to establish AMA1 recombinant adenovirus of Neospora caninum and Toxoplasma gondii, and analysis the immunogenicity of it. According to the open reading frame of N. caninum and T. gondii AMA1 gene sequences, cross universal primers were designed. Based on p MD18T-Nc AMA1 and p MD18T-Tg AMA1 cloning plasmid, recombinant adenovirus shuttle plasmid ADV4-Nc/Tg AMA1 was constructed. Then, ADV4-Nc/Tg AMA1 and pac Ad5 backbone plasmid were linearized and co-transfected 293 T cells. After packaging recombinant adenovirus and measuring the virus titer, collected virus was inoculated into BALB/c mice, confirmed the Ig G antibody levels by indirect ELISA method. The results showed, Nc/Tg AMA1 gene was expressed in Ad5-Nc/Tg AMA1 recombinant adenovirus, Ad5-Nc/Tg AMA1 recombinant adenovirus titer was 109 PFU / mL. Ig G antibody levels in the Ad5-Nc/Tg AMA1 vaccinated inoculating group was significantly higher than p VAX1-Nc/Tg AMA1 plasmid group(P<0.01) and PBS control group(P<0.01). This result indicated that the constructed Ad5-Nc/Tg AMA1 recombinant adenovirus could induce specific humoral immune response in mice, this research laid a solid foundation for the development of a recombinant adenovirus vaccine against N. caninum and T. gondii.
In order to establish AMA1 recombinant adenovirus of Neospora caninum and Toxoplasma gondii, and analysis the immunogenicity of it. According to the open reading frame of N. caninum and T. gondii AMA1 gene sequences, cross universal primers were designed. Based on p MD18T-Nc AMA1 and p MD18T-Tg AMA1 cloning plasmid, recombinant adenovirus shuttle plasmid ADV4-Nc/Tg AMA1 was constructed. Then, ADV4-Nc/Tg AMA1 and pac Ad5 backbone plasmid were linearized and co-transfected 293 T cells. After packaging recombinant adenovirus and measuring the virus titer, collected virus was inoculated into BALB/c mice, confirmed the Ig G antibody levels by indirect ELISA method. The results showed, Nc/Tg AMA1 gene was expressed in Ad5-Nc/Tg AMA1 recombinant adenovirus, Ad5-Nc/Tg AMA1 recombinant adenovirus titer was 109 PFU / mL. Ig G antibody levels in the Ad5-Nc/Tg AMA1 vaccinated inoculating group was significantly higher than p VAX1-Nc/Tg AMA1 plasmid group(P<0.01) and PBS control group(P<0.01). This result indicated that the constructed Ad5-Nc/Tg AMA1 recombinant adenovirus could induce specific humoral immune response in mice, this research laid a solid foundation for the development of a recombinant adenovirus vaccine against N. caninum and T. gondii.
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