口服pCDNA3.1+/Ag85A DNA疫苗在小鼠肠道的表达  

Expression of Oral pCDNA3.1+/Ag85A DNA vaccine in the intestine of mice

作　　者：徐佳[1] 刘滢[2] 于淼[2] 冯永辉[2] 吕昌龙[2] XU Jia LIU Ying YU Miao FENG Yong-hui LYU Chang-long(Department of Pathogenic Biology,Shenyang Medical College in Liaoning Province,Shenyang 110034,China Department of Immunology,China Medical University in Liaoning Province,Shenyang 110122,China)

机构地区：[1]沈阳医学院病原生物学教研室,辽宁沈阳110034 [2]中国医科大学免疫学教研室,辽宁沈阳110122

出　　处：《中国当代医药》2017年第18期4-7,共4页China Modern Medicine

基　　金：沈阳医学院科技基金项目(20132029)

摘　　要：目的分析口服pCDNA3.1+/Ag85A DNA疫苗在小鼠肠道局部的表达情况。方法用脂质体包裹pCDNA3.1+/Ag85A DNA质粒制成口服疫苗。对照组雌性C57BL/6小鼠5只用生理盐水灌胃,实验组雌性C57BL/6小鼠5只用脂质体包裹的Ag85A DNA疫苗灌胃。共免疫3次,每次间隔14 d。免疫结束后14 d处死小鼠,取小肠组织固定于4%多聚甲醛中备用。采用免疫荧光和免疫组化方法检测两组小鼠小肠派氏淋巴结、派氏淋巴结内的树突状细胞和小肠黏膜上皮细胞中Ag85A的表达情况。结果在实验组小鼠的小肠黏膜上皮细胞、派氏淋巴结以及派氏淋巴结内的树突状细胞中均检测到重组pCDNA3.1+/Ag85A DNA疫苗的表达,而且不同部位的小肠黏膜上皮细胞中Ag85A的表达强度不同,靠近固有层的小肠黏膜上皮细胞内Ag85A的表达强度比靠近肠腔侧的小肠黏膜上皮细胞高(P<0.05)。在对照组小鼠上述部位无重组DNA疫苗的表达。结论脂质体包裹的口服DNA疫苗能够被小鼠肠道吸收,同时可诱导特异性黏膜免疫反应。Objective To analyze the expression of oral pCDNA3, l＋/Ag85A DNA vaccine in the intestinal tract of mice. Methods Oral vaccine was made using liposomes to parcel pCDNA3.1 ＋/Ag85A DNA plasmid.5 female C57BL/6 mice in control group were given saline water for gavage while 5 female C57BL/6 mice in experimental group were given li- posome encapsulated Ag85A DNA vaccine for garage.The mice were immunized three times with 14 days interval be- tween each time.In the fourteenth day after immunization,cervical dislocation executed in mice and the small intestine was fixed with 4% paraformaldehyde to be prepared to detect the expression of Ag85A in the intestinal Peyer patches, the dendritic cells in Peyer patches and intestinal epithelial cells with immunofluorescence and immunohistochemistry methods.Results Oral vaccine pCDNA3.1＋/Ag85A DNA was expressed in intestinal Peyer patches,the dendritic cells in Peyer patches and the intestinal epithelial cells of mice in the experimental group,and in different parts of the intestinal epithelial cells in mice of the experimental group,the expression intensity of Ag85A was different.The expression inten- sity of Ag85A was higher near the lamina propria than near the lumen of intestinal epithelial cells （P〈0.05）.h was not expressed in above parts of mice in the control group.Conclusion Oral DNA vaccine parceled with liposomes can be absorbed by intestinal tract of mice and induce specific mucosal immune response.

关 键 词：AG85A DNA疫苗 脂质体 上皮细胞 树突状细胞 小鼠 

分 类 号：R332[医药卫生—人体生理学]