防龋基因疫苗pVAX1-SPG在BALB/c小鼠免疫部位的原位表达  

The expression of anti-caries gene vaccine pVAX1-SPG in BALB/c mice in situ

作　　者：孙天瞳[1] 刘建国[1,2] 姜晶[1] 王伟[1] 亓鹏[1] 白国辉[1,2] 

机构地区：[1]遵义医学院口腔学院,贵州遵义563099 [2]贵州省高等学校口腔疾病研究特色重点实验室暨遵义市口腔疾病研究重点实验室,贵州遵义563099

出　　处：《遵义医学院学报》2015年第6期581-583,590,共4页Journal of Zunyi Medical University

基　　金：贵州省优秀青年科技人才培养计划项目(NO:黔科合人字[2005]0509);贵州省科技创新人才团队建设项目(NO:黔科合人才团队[2013]4026);省市科技合作专项资金项目(NO:省市科合[2014]41);贵州省高等学校特色重点实验室建设项目(NO:黔教合KY字[2013]109)

摘　　要：目的观察防龋基因疫苗p VAX1-SPG免疫BALB/c小鼠后的原位表达。方法重组质粒p VAX1-SPG经颌下腺区皮下注射、股四头肌注射和鼻腔黏膜滴注免疫BALB/c小鼠,免疫组化技术观察目的蛋白Spa P-P/Gbp A-GBD在免疫部位的表达情况。结果小鼠双侧颌下腺组织内均可见目的蛋白的弥漫性表达,尤以导管区域表达强烈,导管细胞的胞浆内呈强阳性染色;股四头肌肌纤维胞浆和肌膜下方阳性染色,呈不均匀的受限表达;鼻腔黏膜上皮下方的固有层与结缔组织中可见呈浅棕黄色染色的目的蛋白表达。结论重组质粒p VAX1-SPG能够在BALB/c小鼠的免疫部位正确表达目的蛋白。Objective To observe the expression of the eukaryotic plasmid p VAX1-SPG as anti-caries gene vaccine,including P domain（Spa P-P）of surface protein antigen（Spa P）and glucan binding domain（GBD）of glucan binding protein A（Gbp A）of streptococcus mutans in situs.Methods BALB/c mice were immunized with plasmid p VAX1-SPG by three different ways including the intranasal immunization,the quadriceps injection and the submandibular gland-targeted injection.Immunohistochemistry technique was used to detect the expression of recombinant protein Spa P-P/Gbp A-GBD.Results The diffused expression of target protein was detected in both sides of submandibular glands,particularly in ducts area with strong positive stain in cytoplasm of duct cells;positive stain in cytoplasm of muscle fiber and under the sarcolemma,but the expression was uneven.The positive light brown stain can be observed in lamina propria and connective tissue of nasal mucosa.Conclusion The fusion protein Spa P-P/Gbp A-GBD can be correctly expressed in vivo.

关 键 词：基因疫苗 变异链球菌 龋齿 表面蛋白 葡聚糖结合蛋白 

分 类 号：R781.1[医药卫生—口腔医学]