摘要
目的观察水凝胶型防龋基因疫苗pVAX1-gtfB/CAT经黏膜途径免疫兔的免疫效果及其目的蛋白在免疫部位的表达。方法 20只新西兰大白兔随机分为5组,每组4只。A组:pVAX1-gtfB/CAT口服组,B组:pVAX1-gtfB/CAT鼻腔黏膜滴注组,C组:空载体pVAX1质粒口服组,D组:空载体pVAX1+水凝胶口服组,E组:生理盐水鼻腔黏膜滴注组。每间隔1周免疫1次,共免疫3次。每次免疫剂量为200μg/只。于免疫前和免疫后第1、2、3、4、6、8、10、12、14、16周采集血液及唾液标本,ELISA法检测血清及唾液中特异性抗体水平。第3次免疫后每组处死1只动物,取免疫部位组织,免疫组织化学检测目的蛋白表达。结果 1水凝胶型防龋基因疫苗pVAX1-gtfB/CAT免疫兔后,诱导了特异性IgG和SIgA抗体产生,在第6和第8周口服免疫诱导的特异性IgG抗体水平高于鼻腔黏膜滴注,其余时间点两种黏膜途径诱导的SIgA和IgG抗体水平差异无统计学意义;2在小肠黏膜和鼻黏膜均检查到了目的蛋白的表达。结论水凝胶型防龋基因疫苗pVAX1-gtfB/CAT经黏膜途径免疫后,目的蛋白能够在免疫部位表达,均能有效诱导兔的系统免疫和黏膜免疫应答。
Objective To observe the immune effect of hydrogel carried anti-caries gene vaccine pVAX1-gtfB /CAT on rabbits by mucosa routes,and investigate the expression of target protein in situ.Methods Twenty New Zealand rabbits were randomly divided into five groups,4 in each group.Group A were immunized with pVAX1-gtfB /CAT by oral immunization,Group B were immunized with pVAX1-gtfB /CAT by intranasal immunization,Group C were immunized with plasmid PVAX1 by oral immunization,Group D were immunized with plasmid PVAX1 combined biodegradable hydrogel by oral immunization and Group E were immunized with normal saline by intranasal immunization,respectively.All the rabbits were taken a total of 3 immunization by receiving 1 immunization every 1 weeks,and every rabbit was immunized at the dose of 200 μg every time.Serum samples and saliva samples were collected before immunization,and at 1,2,3,4,6,8,10,12,14,16 weeks after immunization,respectively.The level of specific antibodies SIgA and IgG in saliva and serum 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 situ by immunohistochemistry,respectively.Results1） The specific antibodies of SIgA and IgG in saliva and serum were significantly increased after immunization in rabbits.The level of antibodies were significantly higher in experimental groups than those in the negative control groups（ P 0.05）.The oral immunization could be superior to other ways at 6 and 8 weeks,but in most of the time the antibodies level were no statistically significant difference.2） Recombinant protein could be detected in the intestine and nasal mucosa.Conclusion Hydrogel carried anti-caries gene vaccine pVAX1-gtfB /CAT on rabbits by mucosa routes can markedly increase the level of specific antibody,and immune responses both in systemic immunity and in mucosal immune system in rabbits.In addition recombinamt protein coucd be correctly expressed in vivo.
出处
《遵义医学院学报》
2014年第5期479-483,共5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)
贵州省高等学校重点学科建设项目(NO:SZXK-[2012]07-04)
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)
关键词
基因疫苗
龋病
水凝胶
免疫
gene vaccine
dental caries
hydrogel
immunization
