中试防龋DNA疫苗pGJA-P/VAX免疫小鼠的实验研究  

Experimental Study of the Pilot-scale Products of Anti-caries DNA Vaccine pGJA-P/VAX in Mice

作　　者：杨亚萍[1] 闵志鹏[1] 樊明文[1] 张爱华[2] 闭兰[2] 黄丽[1] 苑艺芳[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,湖北武汉430079 [2]武汉生物制品研究所

出　　处：《口腔医学研究》2010年第2期157-160,共4页Journal of Oral Science Research

基　　金：国家科技支撑计划项目(编号:2007BAI28B04)

摘　　要：目的:评价中试防龋DNA疫苗pGJA-P/VAX诱导小鼠体内特异性免疫应答的效果。方法:分别由武汉生物制品研究所中试制备和实验室采用德国Qiagen公司去内毒素质粒大量提取试剂盒手工提取防龋DNA疫苗pGJA-P/VAX;两种方法制备的质粒分别于0周,2周经肌肉、鼻腔免疫小鼠,并以鞭毛蛋白作为黏膜佐剂与质粒同时经鼻腔免疫小鼠,PBS及空载体pVAX1作为对照。免疫前开始每两周收集小鼠血清和唾液样本,ELISA定量检测血清及唾液中特异性抗体含量。结果:中试质粒和手提质粒肌注时均能够诱导血清特异性IgG,但诱导唾液特异性sIgA的能力较为有限。在与黏膜佐剂鞭毛蛋白同时滴鼻时,中试质粒可诱导较为明显的血清特异性IgG和唾液特异性sIgA。在相同的免疫条件下,中试质粒诱导的特异性抗体水平低于手提质粒组,但二者无显著性差异。结论:中试防龋DNA疫苗pGJA-P/VAX能够诱导小鼠体内特异性体液免疫应答。Objective： To evaluate the effect of the pilot-scale products of anti-caries DNA vaccine pGJA-P/VAX on inducing specific humoral immune response in mice.Methods： Anti-caries DNA vaccine pGJA-P/VAX were prepared by pilot-scale production in Wuhan Institute of Biologic Products,or extracted and purified using EndoFree Plasmid Maxi Kit in laboratory.Balb/c mice were immunized by both plasmids at week 0 and 2 by intramuscular injection or intranasal immunization.Flagellum protein was used as mucosal adjuvant,and pVAX1 was taken as control.Samples of serum and saliva were collected biweekly.The specific antibody levels were detected by quantitative ELISA.Results： Both the pilot-scale and laboratory products were able to induce specific antibody in serum in groups of intramuscular injection,but had limitations in inducing notable specific sIgA in saliva.While immunized together with flagellum protein,the pilot-scale plasmid was able to induce specific IgG in serum and sIgA in saliva.Compared to laboratory products under the same immunizing condition,the pilot-scale products induced weaker immune response,but with no significant differences.Conclusion： The pilot-scale products of anti-caries DNA vaccine pGJA-P/VAX was able to induce effective humoral immune response.

关 键 词：防龋DNA疫苗 中试 特异性抗体 

分 类 号：R392[医药卫生—免疫学]