日本血吸虫表位肽-DNA颗粒性疫苗对小鼠免疫应答的影响  

Effect of Epitope-based Peptide-DNA Dual Vaccines against Schistosoma japonicum in Mice

作　　者：汪雪峰[1] 张荣波[1] 胡友莹[1] 杜久伟[1] 陈晓军[2] 徐志鹏[2] 苏川[2] 

机构地区：[1]安徽理工大学医学院病原生物学教研室,淮南232001 [2]南京医科大学病原生物学系,江苏省病原生物学重点实验室,南京210029

出　　处：《中国寄生虫学与寄生虫病杂志》2011年第4期319-320,F0003,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No30801046);安徽理工大学硕博基金资助项目(No2010YB004)~~

摘　　要：采用已构建的日本血吸虫Si22．6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（T-PDDV）、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100μlPDDV（含10μgDNA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。EUSA结果显示，T组小鼠脾细胞中IFN-γ的含量[（76．0±11．2）ps／ml]，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。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（10 μg DNA＋28 μg peptide）. One week after the last immunization, the mice were sacrificed, the spleens were removed and splenoeytes were collected. Splenoeyte proliferation was assayed by[3H] TdR incorporation after stimulation with soluble worm antigen （SWA）. Levels of IFN-γ and IL-4 in the splenocyte culture supernatants were determined by ELISA. The results showed that IFN-γ content 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.

关 键 词：日本血吸虫 颗粒性肽-DNA混合疫苗 小鼠 免疫应答 

分 类 号：R383.24[医药卫生—医学寄生虫学]