摘要
目的 本项研究将已构建的编码pac结构基因A P片段的重组质粒pCIA P免疫Wistar大鼠 ,观察重组表面蛋白抗原PAc在大鼠体内不同组织中的原位表达以及免疫定菌鼠后的防龋效果。方法 重组质粒pCIA P经股四头肌肌肉注射和颌下腺区皮下注射两种途径免疫大鼠 ,以免疫组化技术观察重组蛋白PAc在免疫部位的表达。采用股四头肌肌肉注射、颌下腺区皮下注射和颊粘膜下注射 3种方法免疫定菌鼠 ,ELISA法检测血清和唾液中特异性抗体水平 ,Keyes计分法评估免疫定菌鼠后鼠磨牙的患龋情况。结果 大鼠股四头肌细胞中可见PAc蛋白不均匀的受限表达 ,双侧颌下腺组织均可检测到PAc蛋白的阳性染色 ,在导管内表达呈强阳性。用重组质粒pCIA P经颌下腺区皮下注射和颊粘膜免疫的方法可显著增加唾液中抗PAc IgA水平和血清中特异性抗PAc IgG水平。重组质粒免疫定菌鼠后能显著降低定菌鼠龋损计分。特别是颌下腺区皮下和颊粘膜注射免疫组 ,大鼠牙本质龋的破坏程度明显低于其他组。结论 重组质粒pCIA P是一种有效的免疫原 ,粘膜免疫是较理想的DNA防龋疫苗的接种途径。
Objective Protein of Streptococcus mutans is considered as one of the virulence factors due to its ability to mediate the initial attachment of Streptococcus mutans to tooth surface. In this study, an anticaries DNA vaccine pCIA P was used to immunize rats. The expression of PAc in different tissues in vivo, specific immune response and protection effects against dental caries were observed.Methods Plasmid pCIA P was injected into rats by two different routs: intramuscular injection (i.m.) and targeted salivary gland immunization (TSG). Immunohistochemistry technique was used to detect the expression of PAc. Gnotobiotic rats were vaccinated with pCIA P by three different approaches:TSG, intramuscular injection and buccal mucosal injection (i.o.). The specific immune responses were evaluated by ELISA and their anticaries effects were evaluated by Keyes caries scores.Results PAc was expressed in the sarcoplasm and sarcolemma of muscle fibers and submandibular glands, especially strongly positive in duct regions. The levels of serum specific anti PAc IgG and salivary specific anti PAc IgA in TSG immunization and buccal mucosal immunization group were significantly higher than those of other groups. The Keyes caries scores of those two groups were significantly lowere than those of other groups.Conclusion The plasmid pCIA P could provoke specific immune responses as a novel immunogen. Mucosal immunization with pCIA P appears to be an effective genetic immunization method against dental caries
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2002年第1期4-7,T001,共5页
Chinese Journal of Stomatology
基金
国家自然科学基金资助项目 ( 39770 799)
