中国人兽共患病学报2012，Vol.28Issue(10)：1007-1011,5.DOI:10.3969/cjz.j.issn.1002-2694.2012.10.010
幽门螺杆菌hpaA基因在乳酸菌中的表达及免疫原性分析
Expression of Helicobacter pylori hpaA gene in Lactococcus lactis and its immunogenicity analysis
孙振璐 1宫连凤 1刘娟 1王志昱 1高巧 1彭世泽 2韩文清1
作者信息
- 1. 烟台市疾病预防控制中心,烟台,264003
- 2. 北京协和医学院,北京,100005
- 折叠
摘要
Abstract
To develop an oral vaccine against Helicobacter pylori (H. pylori) infection, the H. pylori hpaA gene was expressed in a live delivery vehicle, Lactococcus lactis (L. lactis). The immunogenicity of the recombinant hpaA was evaluated after the L. lactis was administrated orally into mice. Firstly, the hpaA gene of H. pylori was amplified by PCR and cloned in to the prokaryotic expressive vector pNICE: sec. Secondly, the recombinant vector pNICE:sec hpaA was transformated into L. lactis strain NZ9000 to express hpaA protein. Then the recombinant hpaA was induced to express and was identified by SDS PAGE and Western blot. Finally, ICR mice were randomly divided into 4 groups and administrated orally with PBS, L. lactis pNICE:sec, L. lactis pNICE:sec hpaA, and the inactivated H. pylori respectively. The specific IgG and IgA were identified after 7 times immunization. Results showed that the hpaA was amplified and cloned into the vector pNICE:sec sue cessfully. The fusion protein (29 kDa) was expressed in L. lactis by the induction of the nisin. The quantity of expression ac counted for 9. 3 % of the total bacterial protein. Western bolt analysis confirmed that fusion protein could be recognized specif ically by the serum of anti H. pylori. Anti hpaA IgG titers in the serum of L. lactis pNICE:sec hpaA group was higher than that in the L. lactis pNICE:sec group and no obviously difference with the inactivated H. pylori. The anti hpaA IgA titer in the intestinal mucosa of L. lactis pNICE: sec hpaA group was higher than that in other groups. The results suggest that the re combinant L. lactis which expresses hpaA, might be applicable as an oral vaccine to induce protective immunity against H. pylori.关键词
幽门螺杆菌/乳酸乳球菌/hpaA基因Key words
Helicobacter pylori/ Lactococcus lactis / hpaA gene分类
医药卫生引用本文复制引用
孙振璐,宫连凤,刘娟,王志昱,高巧,彭世泽,韩文清..幽门螺杆菌hpaA基因在乳酸菌中的表达及免疫原性分析[J].中国人兽共患病学报,2012,28(10):1007-1011,5.
