变形链球菌防龋基因疫苗pcDNA3-gtfB免疫途径筛选的实验研究

A Study on Screening Effective Immunization Route of Anticaries DNA Vaccine pcDNA3-gtfB

目的 :利用含有葡糖基转移酶抗原基因的重组质粒pcDNA3_gtfB作为基因疫苗 ,筛选有效的免疫途径。方法 :重组质粒pcDNA3_gtfB通过股四头肌注射、鼻腔灌注和颌下腺周注射免疫Wistar大鼠 ,采用ELISA法测定血清IgG、唾液IgA的动态变化。结果 :经股四头肌注射免疫后产生的血清IgG抗体水平明显高于其它两组血清IgG抗体水平 (P <0 0 1) ,且鼻腔灌注组和颌下腺周注射组间的血清IgG抗体水平无显著性差异 (P >0 0 5 )。各组唾液S_IgA抗体水平均存在显著差异 (P <0 0 1) ,腺周注射组产生的唾液S_IgA抗体水平最高 (P <0 0 1)。结论 :基因疫苗通过腺周注射免疫途径能最有效激发特异性唾液S_IgA抗体的产生 ,可望成为一种有效的防龋基因疫苗免疫途径 。

Objective:Glucosyltransferase-B (GTF-B) of Streptococcus mutans has been implicated as a principal virulent factor in the development of dental caries. The objective was to use recombined plasmid pcDNA-gtfB expressing multiple antigen of glucosyltransferase-B as gene vaccine to immunize rats through different route, and to investigate the immunization effects of immunization routes.Methods: A total of 18 Wistar rats were divided into 3 groups, including the quadriceps injection group, the intranasal irrigation group and the submandibular gland-targeted injection group. The serum IgG and salivary IgA were assayed by using ELISA after pcDNA3-gtfB immunization. The serum IgG and salivary IgA in different groups were compared using statistical one-way ANOVA.Results: Compared these 3 groups, the serum IgG in the quadriceps injection group was much higher than those of other two groups (P<0.01), while the salivary IgA of the submandibular gland-targeted injection was much higher than those of other two groups (P<0.01).Conclusion: It is indicated pcDNA3-gtfB is good candidate for anticarious gene vaccine, and submandibular gland-targeted injection is an effective immunization route for stimulating salivary IgA.