日本血吸虫重组28GST T细胞表位谱的预测及鉴定  

Prediction of T-cell epitopes from schistosoma japonicum 28 kDa glutathione-S-transferase and identification of their Th1 type T-cell epitopes

作　　者：李光富[1] 张兆松[1] 王勇[1] 王新军[1] 朱翔[1] 季旻珺[1] 吴观陵[1] 

机构地区：[1]南京医科大学病原生物学系,江苏南京210029

出　　处：《细胞与分子免疫学杂志》2004年第3期352-355,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学资金资助项目 (No .30 2 71 1 66)

摘　　要：目的 :用软件预测日本血吸虫重组 2 8GST抗原分子T细胞表位谱 ,并鉴定其Th1型细胞表位。方法 :用软件预测重组2 8GSTT细胞表位谱 ,并筛选几种较好的T细胞表位 ,人工合成表位肽或用基因工程制备重组表位肽融合蛋白。体外刺激经照射的尾蚴感染并用 2 8GST加强免疫的C5 7BL/ 6小鼠 (H 2 b)脾细胞 ,通过淋巴细胞增殖试验、ELISA及流式细胞术等 ,分析各种表位的免疫刺激作用 ,鉴定Th1型细胞表位。结果 :在 9个候选的表位中 ,P6 (73～ 86aa)是刺激作用最强的Th1型细胞表位。结论 :重组 2AIM: To predict T cell epitopes of recombinant schistosoma japonicum 28 kDa glutathione S transferase(GST) with softwares and identify the Th1 type T cell epitopes by experiments. METHODS: T cell epitopes of recombinant schistosoma japonicum 28kDa GST were predicted with softwares and several epitopic candidates were screened from them according to their scores. Some of the epitopic candidates were synthesized and the other epitope peptides fused with thioredoxin(Trx) were expressed in E.coli BL21(DE3) and purified by Ni + column affinity chromotography. C57BL/6 (H 2 b) mice’s were immunized via peritoneal infection with ultraviolet ray irradiated cercariae and then boosted with recombinant schistosoma japonicum 28 kDa GST. The immunized mice splenocytes were prepared, cultured and stimulated with synthesized epitope peptides and epitope peptide fusion proteins, respectively. Stimulation activity of synthesized epitope peptides and epitope peptides fusion proteins were assayed by lymphocyte proliferation assay. Levels of IFN γ and IL 2 were measured by ELISA. CD4 + T cells and T cells secerting IFN γ and IL 4 were detected by flow cytometry. RESULTS: Epitope P6(73 86aa) among 9 epitopic candidates could generate the strongest stimulation effect on splenocytes, stimulate secretion of higher levels of IFN γ and IL 2, and induce more IFN γ + and IL 4 + T cells. CONCLUSION: The recombinant 28 kDa GST possesses functional Th1 type T cell epitope.

关 键 词：日本血吸虫 表位 谷胱甘肽-S-转移酶(GsT) 

分 类 号：R383.24[医药卫生—医学寄生虫学]