复苏因子E-DNA疫苗对BALB/C小鼠免疫效应的研究  

The immunological effects of Rpf-E-DNA on BALB/C mice

作　　者：邢爱英[1] 刘忠泉[1] 贾红彦[1] 李自慧[1] 郑晓静[1] 曹廷明[1] 杜风娇[1] 杜博平 古淑香[1] 张宗德[1] 

机构地区：[1]北京市结核病胸部肿瘤研究所结核病分子生物学研究室,101149

出　　处：《实用医学杂志》2010年第23期4259-4261,共3页The Journal of Practical Medicine

基　　金：国家"十一五"重大科技专项(编号:2008ZX10003-001)

摘　　要：目的:探索复苏因子E(RpfE)-DNA疫苗的小鼠免疫效果。方法:利用真核表达载体PCDNA3.1构建PCDNA3.1-RpfE重组质粒。将SPF级BALB/C小鼠随机分为5组,即空质粒组、RpfE质粒组、BCG组、生理盐水组、空白对照组,于0、3、6周先后3次免疫小鼠,在第3次免疫后3周,心脏取血ELISA方法检测血清RpfE IgG抗体效价、干扰素γ(IFNγ)水平;并行脾淋巴细胞培养,以RpfE蛋白刺激淋巴细胞,CCK-8法检测淋巴细胞增殖指数、ELISA方法检测上清诱导IFNγ和白介素12(IL-12)产生水平,乳酸脱氢酶试验检测小鼠脾细胞毒性T淋巴(CTL)杀伤效应。结果:(1)RpfE质粒组小鼠血清中RpfE抗体水平明显高于其他各组(P<0.001);RpfE质粒组IFNγ水平明显高于空质粒组、生理盐水组、空白对照组和BCG组(P<0.01或P<0.05)。(2)RpfE质粒组具有较强的刺激淋巴细胞增殖作用,CCK-8实验表明其增殖指数高于其他各组(P<0.05);上清ELISA检测表明IFNγ和IL-12产生水平也高于其他各组(P<0.05)。(3)RpfE质粒组脾淋巴细胞特异性CTL杀伤性与其他各组比较有统计学意义(P<0.05)。结论:RpfE-DNA疫苗可诱导小鼠的免疫应答,具有良好的体液和细胞免疫原性。Objective To explore the immunological effects of resuscitation-promoting factor E （RpfE）-DNA on BALB/C mice. Methods Plasmid PCDNA3.1-RpfE was constructed by eukaryotic expression vector PCDNA3.1. SPF-BALB/C mice were randomly divided into 5 groups：empty plasmid group,RpfE vaccine group,BCG group,normal saline group,and blank control group. The mice were immunized on weeks 0,3,and 6. Three weeks after the third immunization,blood was collected from the heart to detect serum IgG antibody to RpfE and IFNγ by ELISA; spleen lymphcytes were cultured and then stimulated with Rpf E protein lymphocyte proliferation index was detected with CCK-8 technique,so were levels of IFNγ and IL-12 induced in the supernatant by ELISA; the killing effect of CTLs was determined by LDH assay. Results Levels of serum IgG antibody to RpfE was significantly higher in RpfE vaccine group than in other groups （P〈0.001）. Rpf E vaccine had a stimulating effect on lymphocyte proliferation,with a higher proliferation index than other groups（P〈0.05）. Levels of IFNγ and IL-12 were also higher in RpfE vaccine group（P〈0.05）. As compared with other groups,the specific CTLs in RpfE vaccine group had a greater killing effect（P〈0.05）. Conclusions RpfE-DNA vaccine can induce immune responses in BALB/C mice,with good humoral and cellular immunogenicity.

关 键 词：疫苗 DNA RpfE 免疫 

分 类 号：R392[医药卫生—免疫学]