【摘要】 采用已构建的日本血吸虫Sj22.6抗原CTL、Th和B细胞表位肽-DNA颗粒性疫苗（PDDV）及其混合疫苗免疫C57BL/6小鼠。36只小鼠随机均分6组,即18K对照组([K]₁₈-空质粒PDDV)、PBS对照组、C组（C-PDDV）、T组（TPDDV）、B组（B-PDDV）和C-T-B组（C-PDDV、T-PDDV和B-PDDV等量混合）,每鼠分别在第0、3和6周麻醉下经尾脊部皮下注射100μl PDDV（含10μg DNA和28μg肽）,对照组注射等量的空质粒DNA和[K]₁₈肽或PBS。末次免疫后7d,脱颈处死,制备脾细胞悬液,经日本血吸虫成虫抗原（SWA）刺激后根据³H标记的胸腺嘧啶核苷（³H-TdR）检测脾细胞增殖反应,ELISA法检测脾细胞培养上清中γ干扰素（IFN-γ）和白细胞介素-4（IL-4）的含量。ELISA结果显示,T组小鼠脾细胞中IFN-γ的含量[（76.0±11.2）pg/m1],高于PBS[（13.0±2.1）pg/ml]和18K对照组[（14.0±3.2）pg/ml]（P<0.01）,T组和C-T-B组小鼠脾细胞中IL-4的水平分别为（152.0±21.1）和（86.0±12.2）pg/ml,高于其他组（P<0.01或P<0.05）。T组小鼠脾细胞经SWA刺激后,增殖反应明显高于PBS和18K对照组（P<0.01）;而C-T-B组小鼠脾细胞的增殖反应与PBS和18K对照组相比,差异无统计学意义（P>0.05）。说明T-PDDV和C-T-B混合PDDV诱导的免疫应答强于单价的C-PDDV和B-PDDV。更多还原

【Abstract】 A C-T-B PDDV mixture of the three constructed epitope-based peptide-DNA dual vaccines(PDDV) containing the CTL(C),Th(T) and B-cell(B) epitopes from Sj22.6 tegument(C-PDDV,T-PDDV and B-PDDV) with a 1:1:1 ratio was prepared.Thirty-six mice were randomly divided into six groups averagely named as 18K group,PBS group,C-PDDV group,T-PDDV group,B-PDDV group,and C-T-B PDDV group.All the mice received three immunizations at 2-week intervals with the same dose of antigen(10μg DNA+28μg peptide).One week after the last immunization,the mice were sacrificed,the spleens were removed and splenocytes were collected.Splenocyte proliferation was assayed by[~3H]TdR incorporation after stimulation with soluble worm antigen(SWA).Levels of IFN-γand IL-4 in the splenocyte culture supematants were determined by ELISA.The results showed that IFN-γcontent in T-PDDV group[(76.0±11.2) pg/ml]was higher than that of PBS[(13.0±2.1) pg/ml]and 18K control groups[(14.0±3.2) pg/ml](P<0.01).IL-4 level in T-PDDV [(152.0±21.1) pg/ml]and C-T-B mixture groups[(86.0±12.2) pg/ml]was higher than others(P<0.01 and P<0.05).The splenocytes from T-PDDV group showed a significant increase in proliferation compared with PBS and 18K control groups after stimulation by SWA(P<0.01).However,there was no significant difference in splenocyte proliferation among C-T-B, PBS and 18K control groups(P>0.05).These findings indicate that T-PDDV and C-T-B PDDV mixture induces stronger immune response than that of C-PDDV or B-PDDV.更多还原