沙眼衣原体主要外膜蛋白多表位重组DNA诱导小鼠体液免疫有效剂量的探讨  

Evaluation of the optimum immune dose for a DNA vaccine in mice based on the major outer membrane multiepitope from Chlamydia trachomatis

作　　者：饶品欢[1] 石朝晖[2] 李文姝[1] 吕艳[1] 陈向敏[1] 朱珊丽[1] 张丽芳[1] 

机构地区：[1]温州医学院微生物学与免疫学教研室分子病毒与免疫研究所,浙江温州325035 [2]平顶山市疾病预防控制中心检验科,河南平顶山467000

出　　处：《温州医学院学报》2012年第2期103-106,共4页Journal of Wenzhou Medical College

基　　金：国家自然科学基金资助项目(30972669);浙江省自然科学基金资助项目(Y2100611)

摘　　要：目的:探讨沙眼衣原体主要外膜蛋白多表位(Ct MOMP168)重组真核表达质粒pcDNA3.1(+)/Ct MOMP168免疫小鼠的有效免疫剂量,为体内免疫学效应的研究提供实验依据。方法:在重组真核表达质粒pcDNA3.1(+)/Ct MOMP168构建成功的基础上,抽提纯化质粒,分3个免疫剂量组,即50、100和150μg剂量组,同时设PBS对照组。肌肉注射免疫BALB/c小鼠3次,间隔2周,于免疫的0、2、4、6、8周分别尾静脉取血,分离血清,间接ELISA法检测小鼠血清中Ct MOMP多表位特异性抗体IgG的生成量,并于0、1、3、5、7周取小鼠阴道冲洗物,检测Ct MOMP多表位特异性SIgA的生成量,分别进行统计学分析。结果:3个不同免疫剂量组,血清特异性抗体IgG生成量于免疫第4周后均开始随免疫时间延长而逐步增加,150μg剂量组抗体IgG的生成水平高于其他剂量组(P<0.05)。免疫小鼠阴道分泌物中特异性抗体SIgA生成较快,免疫开始后1周,150μg剂量组SIgA的产生水平高于其他组,7周后SIgA开始下降,但150μg剂量组抗体水平仍高于其他剂量组(P<0.05)。结论:重组质粒DNA免疫小鼠的特异性抗体生成显示了剂量依赖关系,高剂量(150μg)免疫组小鼠血清特异性抗体IgG和阴道分泌性特异性抗体SIgA生成水平优势显著。Objective： To evaluate the optimum immune dose for a DNA vaccine of the major outer membrane multiepitope（MOMP168） from Chlamydia trachomatis（Ct） in mice,in order to directly offer valuable experimental information for the following research about immuno-logical effects.Methods： Based on the successful construction and purification of recom-binant plasmid pcDNA3.1（＋）/Ct MOMP168,BALB/c mice were administered respectively with three different immune dose of 50,100 and 150μg of pcDNA3.1（＋）/Ct MOMP168 by intra-muscular injection,and control group was administered with PBS buffer.All mice were repeatedly immunized three times at two weeks intervals.Serum was collected in week 0,2,4,6,8 for measurement of specific antibodies IgG against recombinant Ct MOMP168 protein with ELISA,while,the vaginal washings were collected in week 0,1,3,5,7 to measure sIgA against recombinant Ct MOMP168 protein also with ELISA,and then the data were handled by statistical analysis.Results： The production of specific IgG in serum from immunized mice with three different immunization doses began to gradually increase with the prolonged immunity,and the level of IgG in group mice immunized with maximum dose of 150μg was higher than that in other groups（P 0.05）.While,the production of specific secret IgA in vaginal washings generated fast,in week 1 after immunization,the level of sIgA in group mice immunized with maximum dose of 150μg was higher than that in other groups（P 0.05）,sIgA began to decline in week 7,but the level of sIgA in mice immunized 150μg dose was still higher than that in other groups（P 0.05）.Conclusion： The production of specific antibody in mice administered by DNA vaccine shows a dose-dependent relationship.The mice immunized with the selected immune dose of 150μg generate a higher level of IgG in serum and sIgA in vaginal washings compared with other immune dose.

关 键 词：沙眼衣原体 主要外膜蛋白 DNA免疫 免疫剂量 小鼠 

分 类 号：R374.1[医药卫生—病原生物学]