靶向融合防龋DNA疫苗免疫大鼠的研究  

Immunization of rats with a targeted fusion anticaries DNA vaccine

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

机构地区：[1]武汉大学口腔医学院口腔生物医学工程教育部重点实验室,430079

出　　处：《中华口腔医学杂志》2004年第6期459-462,共4页Chinese Journal of Stomatology

基　　金：国家自然科学基金资助项目 (3 0 2 714 16)

摘　　要：目的　检测靶向融合防龋DNA疫苗 pGJA P免疫大鼠后的原位表达。比较 pGJA P与融合防龋DNA疫苗pGLUA P免疫定菌鼠后产生的抗体水平和防龋效果。方法　质粒pGJA P分别经股四头肌注射和鼻腔滴注免疫大鼠 ,免疫组化法检测重组蛋白在免疫部位的表达。建立定菌鼠模型 ,质粒pGJA P和 pGLUA P分别经股四头肌注射和鼻腔滴注免疫定菌鼠 ,ELISA法检测血清和唾液中的特异性抗体水平 ,取上下颌骨进行Keyes龋齿记分。 结果　在 pGJA P肌肉免疫组的股四头肌和鼻腔免疫组的鼻腔黏膜检测到了表达的重组蛋白。pGJA P肌肉免疫组的血清抗PAc和抗GTFIgG滴度显著高于其他组 (P <0 0 1)。pGJA P肌肉免疫组和 pGJA P鼻腔免疫组的唾液抗PAc和抗GTFIgA滴度显著高于其他组 (P <0 0 1)。pGJA P免疫组的龋齿记分显著低于其他组 (P <0 0 1)。结论　pGJA P在动物体内能够正确表达。与pGLUA P相比 ,pGJA P能诱导更强的体液免疫反应 。Objective To observe the expression of a targeted fusion anticaries DNA vaccine pGJA-P in situ. To compare the levels of specific antibodies and anticaries efficacy generated by pGJA-P and pGLUA-P, a fusion anticaries DNA vaccine.Methods pGJA-P was administrated intramuscularly or intranasally to rats, and the expression of recombinant protein was detected by immunohistochemistry technique. Wistar rats were fed a cariogenic diet and orally infected with S. mutans , then immunized with pGJA-P or pGLUA-P via the intramuscular or intranasal route. All rats received a booster immunization 2 weeks later. At the termination of the experiment, blood and saliva samples were collected for assay of antibodies by ELISA and jaws were obtained for caries evaluation by the Keyes method. Results Recombinant protein could be detected in muscle in intramuscularly immunized rats and in nasal mucosa in intranasally immunized rats. Rats immunized intramuscularly with pGJA-P had significantly higher serum IgG levels than others( P <0 01). Rats immunized intranasally or intramuscularly with pGJA-P had significantly higher salivary IgA levels than others( P <0 01). Keyes scores of pGJA-P groups were significantly lower than those of pGLUA-P groups and pCI groups( P <0 01).Conclusions pGJA-P could be correctly expressed in vivo. pGJA-P generated increased humoral immune response and anticaries efficacy compared with pGLUA-P.

关 键 词：免疫学 大鼠 龋齿 预防 检测 变形链球菌 靶向融合防龋DNA疫苗 

分 类 号：R780.1[医药卫生—口腔医学]