期刊论文

Yu, W.

中文

微生物学报

0001-6209

2010

50

4

554-559

10.13343/j.cnki.wsxb.2010.04.019

湖南省自然科学基金优秀面上项目（06JJ2093） 湖南省教育厅青年基金（08B067）~~

本校为第一机构

[目的]观察pcDNA3.1(+)/Lpp20-IL2免疫C57BL/6小鼠后所产生的体液免疫和细胞免疫应答水平,为研制高效、新型的幽门螺杆菌核酸疫苗提供实验依据.[方法]构建pcDNA3.1(+)/Lpp20-IL2重组载体,并转染HeLa细胞,用Westem-blot观察鉴定其在真核细胞得到表达后免疫C57 BL/6小鼠,ELISA间接法测定小鼠血清中抗Lpp20 IgG抗体水平,ELISA双抗体夹心法检测脾淋巴细胞培养上清中IFN-γ、IIA水平,MTT比色法检测脾淋巴细胞增殖反应,免疫荧光组化法检测Lpp20蛋白在小鼠肌肉组织中的表达情况.[结果]成功构建了pcDNA3.1(+)/Lpp20-IL2真核表达载体,且重组质粒能在HeLa细胞内有效表达目的蛋白;小鼠接种pcDNA3.1(+)/Lpp2...

OBJECTIVES: To evaluate the humoral and cellular immune responses in C57BL/6 mice induced by pcDNA3.1(+)/Lpp20-IL2 for further development of DNA vaccine against H. pylori infection. METHODS: The eukaryotic expression vector pcDNA3.1(+)/Lpp20-IL2 was constructed and then transfected into HeLa cells using liposome. Western blot was used to verify the expression of Lpp20 antigen gene in HeLa cells. Then five groups of 6-week-old C57BL/6 mice were immunized intramuscularly with either of the following vaccines: pcDNA3.1(+)/Lpp20-IL2, pcDNA3.1(+)/Lpp20, pcDNA3.1(+)/IL2, mock plasmid pcDNA3.1(+), o...