靶向融合防龋DNA疫苗经肌肉免疫动物的实验研究  

Immunization with targeted fusion anticaries DNA vaccine via intramuscular route:experiment with murine

作　　者：于飞[1] 樊明文[1] 许庆安[1] 贾荣[1] 郭继华[1] 边专[1] 陈智[1] 彭彬[1] 范兵[1] 

机构地区：[1]武汉大学口腔生物医学工程教育部重点实验室,430079

出　　处：《中华医学杂志》2004年第9期754-759,共6页National Medical Journal of China

基　　金：国家自然科学基金资助项目 (3 0 2 714 16)

摘　　要：目的　观察靶向融合防龋DNA疫苗pGJA P在大鼠肌肉组织中的原位表达。检测靶向融合防龋DNA疫苗pGJA P经股四头肌注射途径免疫大鼠后体内抗体水平和防龋效果 ,并与融合防龋DNA疫苗pGLUA P进行比较。在小鼠体内长期观察抗体动态变化。方法　质粒pGJA P经股四头肌注射途径免疫大鼠 ,免疫组织化学方法检测重组蛋白的原位表达。质粒pGJA P和pGLUA P经股四头肌注射途径免疫定菌鼠 ,ELISA法检测血清IgG和唾液IgA抗体水平 ,Keyes记分法评估定菌鼠磨牙患龋情况。质粒pGJA P和pGLUA P经股四头肌注射途径免疫BALB/c小鼠 ,ELISA法检测血清IgG和唾液IgA动态变化。结果　大鼠股四头肌组织检测到表达的重组蛋白。大鼠pGJA P免疫组血清抗PAcIgG水平 (1∶2 0 0 0 0 0 )和抗GTFIgG水平 (1∶5 80 0 0 )均显著高于pGLUA P免疫组 (1∶2 30 0 0和 1∶110 0 0 )(均P <0 0 1)。大鼠pGJA P免疫组唾液抗PAcIgA水平 (1∶8)和抗GTFIgA水平 (1∶6 )均显著高于pGLUA P免疫组 (1∶2和 1∶2 ) (均P <0 0 1)。pGLUA P免疫组未能引起有效的唾液IgA反应。pGJA P免疫组龋齿记分显著低于pGLUA P免疫组和空白组 (P <0 0 1)。小鼠pGJA P免疫组有效血清IgG、唾液IgA和pGLUA P免疫组有效血清IgG均持续至本实验结束 ,pGJAObjectives To observe the expression of a targeted fusion anticaries DNA vaccine pGJA-P in muscular in vivo. To compare the levels of specific antibodies and anticaries efficacy generated by pGJA-P and fusion anticaries DNA vaccine pGLUA-P in gnotobiotic rats,and observe the kinetics of antibody responses in BALB/c mice. Methods (1) Twelve 28-day-old female Wistar rats were randomly divided into 2 groups of 6 rats to be injected with the plasmid pGJA-P containing the signal peptide and extracellular regions of human CTLA 4,hinge and Fc regions of human IgG,the glu sequence of gtfB gene and A-P fragment of pac gene of Streptococcus mutans or the eukaryotic expression plasmid pCI into the quadriceps muscle of thigh respectively. Three days after the rats were killed and specimens of quadriceps muscles of thigh were taken. Immunohistochemical SP staining was used to examine the in situ expression of pGJA-P. (2) Twenty-four 18-day-old female Wistar rats were randomly divided into 4 groups of 6 rats. The rats were fed with cariogenic food. During the age of 20～22 days cariogentic food containing broad-spectrum antibiotics was fed. Then aseptic cotton stick was used to swab the oral cavity and be smeared onto the solid medium so as to observe the growth of bacteria under anaerobic culture for 48 hours. During the age of 24～26 days,S. mutans Ingbritt cultured anaerobically was swab onto the surface of teeth of the rats twice with an interval of 30 minutes. After the inoculation aseptic cotton stick was used to wipe the oral cavity and be smeared onto the solid medium so as to observe the growth of bacteria under anaerobic culture for 48 hours. When the gnotobiotic rats were 28 days old they were injected with pGJA-P,pGLUA-P,fusion anticaries DNA voccine against both PAc,cell surface proteon antigen,and glucosyltransferase (GTF),pCI or normal saline into the quadriceps muscle of thigh respectively,2 weeks later a booster shot was given. When the rats were 63 days old their saliva and blood samples were collected. Th

关 键 词：靶向融合 防龋DNA疫苗 免疫动物 IGA 链球菌龋齿 

分 类 号：R392.1[医药卫生—免疫学]