CpG寡核苷酸的筛选及其对rSj28GST的免疫佐剂作用  

SELECTION OF CPG ODN AND ITS ADJUVANT EFFECT ON ANTIGEN rSj28GST

作　　者：李光富[1] 张兆松[1] 朱鸿飞[2] 王勇[1] 朱翔[1] 王新军[1] 季旻珺[1] 刘丰[1] 蔡晓萍[1] 吴海玮[1] 吴观陵[1] 

机构地区：[1]南京医科大学病原生物学系,南京210029 [2]南京农业大学生物工程研究所

出　　处：《中国血吸虫病防治杂志》2004年第2期104-108,共5页Chinese Journal of Schistosomiasis Control

基　　金：国家"8 63"高技术研究发展计划项目 ( No.2 0 0 1AA2 43 0 71);国家自然科学基金项目 ( No.3 0 2 71166)

摘　　要：目的　研究 Cp G寡核苷酸 (Cp G ODN)对日本血吸虫重组 2 8k Da GST(r Sj2 8GST)抗原分子的免疫佐剂作用和诱导 Th1型反应的效应。方法　用淋巴细胞增殖试验筛选对鼠有良好免疫刺激作用的 Cp G ODN;用基因工程方法生产 r Sj2 8GST抗原分子 ;EL ISA检测小鼠抗 r Sj2 8GST的同型抗体 Ig G1 、Ig G2 a水平。结果　筛选出的 Cp G ODN如 182 6、2 0 0 2、182 6 4、2 10 2等具有良好的免疫刺激活性 ,其 SI>2 ;纯化的 r Sj2 8GST抗原分子 ,呈单一电泳条带 ;以 Cp G ODN为 r Sj2 8GST免疫佐剂 ,其免疫小鼠产生的抗体滴度显著高于对照组 ,且同型抗体 Ig G2 a与 Ig G1 比值也明显升高。结论　 Cp G ODN182 6 4是一种良好的免疫佐剂 ,并可显著诱导Objective To study the adjuvant effect of CpG ODN on recombinant Schistosoma japonicum 28 kDa glutathione-S-transferase fusion protein(rSj28GST-Trx) and the activity inducing Th1 type response. Methods The stimulation activities of different CpG ODNs on mice splenocytes were assayed in vitro by the lymphocyte proliferation assay. rSj28GST-Trx was expressed in E.coli containing recombinant plasmid Sj28GST/pET32c and purified by washing its inclusion body and Ni + column affinity chromotography. The levels of isotypic antibodies IgG 2a and IgG 1 were measured by ELISA. Results Splenocyte proliferation of C57BL/6 mice was stimulated well by the selected CpG ODNs 1826, 2002, 18264, 2102, with SI>2. The rSj28GST-Trx was a single band identified by SDS-PAGE. The levels of anti-rSj28GST-Trx antibodies and isotypic antibodies IgG 2a and IgG 1 in experimental C57BL/6 mice were higher than those in the control after immunization with rSj28GST-Trx and adjuvant CpG ODN. Conclusion The modified CpG ODN 18264 is a good adjuvant, and can induce significantly Th1 type response.

关 键 词：CPG寡核苷酸 日本血吸虫 谷胱甘肽-S-转移酶 免疫佐剂 

分 类 号：R383.24[医药卫生—医学寄生虫学]