摘要
目的:观察温敏型生物降解水凝胶PLGA-PEG-PLGA搭载防龋基因疫苗pVAX1-spap/A的免疫效果及重组质粒的原位表达情况。方法:28只新西兰大白兔随机分为7组。首次免疫后1周加强免疫,再1周加强免疫第2次,共3次。每次免疫剂量200μg/只。于免疫前1d和免疫后第1,2,3,4,6,8,10,12,14,16周采集动物血液及唾液标本,间接ELISA法检测特异性抗体IgA和IgG。结果:以温敏型生物降解水凝胶PLGA-PEG-PLGA作为载体的防龋基因疫苗pVAX1-spap/A免疫新西兰大白兔后,特异性IgA和IgG抗体水平明显升高,持续14周,与对照组比较差异有统计学意义。在免疫部位的检查到目的蛋白表达。结论:温敏型生物降解水凝胶PLGA-PEG-PLGA作为载体的防龋基因疫苗pVAX1-spap/A经四种途径免疫新西兰大白兔后,目的蛋白能够在免疫部位表达,能够有效诱导动物的体液免疫和黏膜免疫应答。
Objective：To observe the immune effect of temperature-sensitive biodegradable hydrogel carried anti-caries gene vaccine pVAX1-spap/A in rabbits by different routes,and investigate the expression of recombinant plasmid pVAX1-spap/A in vivo.Methods：28New Zealand rabbits were randomly divided into seven groups.All rabbits were taken a total of 3immunization by receiving 1immunization every 1weeks,and every rabbit was immunized at the dose of 200μg every time.Serum samples and saliva samples were collected before immunization,and at1,2,3,4,6,8,10,12,14,16 weeks after immunization,respectively.The level of specific antibodies IgA and IgG in serum and saliva were determined using enzyme-linked immunosorbent assay.After the third immunization,one rabbit of every group were sacrificed to observe the expression of recombinant protein in vivo by immunohistochemistry,respectively.Results：The specific antibodies of IgA and IgG in saliva and serum were significantly increased after immunization in rabbits,and this has continued for more than 14 weeks.The level of antibodies were significantly higher in experimental groups than those in the negative control groups（P〈0.05）.Recombinant protein could be detected in quadriceps,submandibular gland,intestine and nasal mucosa in vivo.Conclusion：Temperature-sensitive biodegradable hydrogel carried anti-caries gene vaccine pVAX1-spap/A by quadriceps injection,oral immunization,submandibular gland injection and intranasal immunization can markedly increase the level of specific antibody,and immune responses both humoral and mucosal in rabbits.In addition,recombinant protein could be correctly expressed in vivo.
出处
《口腔医学研究》
CAS
CSCD
2014年第10期934-938,共5页
Journal of Oral Science Research
基金
贵州省优秀青年科技人才培养计划项目(黔科合人字[2005]0509号)
贵州省科技创新人才团队建设项目(黔科合人才团队[2013]4026)
贵州省高等学校重点学科建设项目(SZXK-201207-04号)
贵州省高等学校特色重点实验室建设项目(黔教合KY字[2013]109)
关键词
基因疫苗
龋病
水凝胶
免疫
Gene vaccine Dental caries Hydrogel Immunization
