幽门螺杆菌靶向核酸疫苗的构建及免疫学评价
幽门螺杆菌;核酸疫苗;靶向免疫;间接免疫荧光;酶联免疫吸附试验,张瑞平,刘纯杰,陶好霞,张兆山,张瑞平,通讯作者:,ConstructionoftargetedDNAvaccineofHpyloriandimmunetestinBALB,cmice
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张瑞平, 刘纯杰, 陶好霞, 张兆山, 军事医学科学院生物工程研究所 北京市 100071
张瑞平, 讲师, 在读博士, 主要从事基因工程疫苗研究.
国家高技术研究发展计划(863)资助项目, No. 2004AA215213
通讯作者: 张兆山, 100071, 北京市丰台区东大街20号, 军事医学科学院生物工程研究所四室. zhangzs@nic.bmi.an.cn
电话: 010-66948834
收稿日期: 2007-01-25 接受日期: 2007-04-02
Construction of targeted DNA vaccine of H pylori and immune test in BALB/c mice
Rui-Ping Zhang, Chun-Jie Liu, Hao-Xia Tao, Zhao-Shan Zhang
Rui-Ping Zhang, Chun-Jie Liu, Hao-Xia Tao, Zhao-Shan Zhang,Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China
Supported bythe Hi-Tech Research and Development Program of China, No. 2004AA215213
Correspondence to:Professor Zhao-Shan Zhang, Beijing Institute of Biotechnology, Academy of Military Medical Sciences, 20 Dongda Street, Fengtai District, Beijing 100071, China. zhangzs@nic.bmi.an.cn
Received:2007-01-25 Accepted:2007-04-02
Abstract
AIM: To construct DNA vaccine targeted on antigen-presenting cells for the purpose of increasing the immunogenecity of Helicobacter pylori DNA vaccine.
METHODS: DNA vaccine was constructed by combining the targeted DNA sequence with katA. Whether or not the DNA vaccine could be expressed in the mammalian cells was detected by indirect immunofluorescence assay of 293T cells transfected with DNA vaccine. The IgG, IgG1 and IgG2a antibody titers of BALB/c mice immunized with DNA vaccine were also determined.
RESULTS: It was indicated in the immunofluorescence assay of 293T cells transfected with DNA vaccine that the KatA protein could be expressed in these cells. Enzyme-linked immunosorbent assay (ELISA) also showed that the transfected cells with pcDNAkathIgz had a higher affinity for IgG. The IgG antibody titer of BALB/c mice immunized with targeted DNA vaccine was significantly higher than that of mice immunized with pcDNAkatA, and a shift form (Th2 response to Th1 response) was achieved in the mice immunized with DNA vaccine ......
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