sLAG-3-Ig的克隆、表达及其生物活性的实验研究  

Cloning,expression and characterization of sLAG-3-Ig

作　　者：新宇[1] 吴昌归[1] 刘颖格[1] 史皆然[1] 陈卫强[1] 赵峰[1] 戴艳丽[1] 

机构地区：[1]第四军医大学西京医院呼吸内科,陕西西安710032

出　　处：《中国病理生理杂志》2006年第5期893-895,共3页Chinese Journal of Pathophysiology

基　　金：陕西省自然科学基金资助项目(No.2002C2-16)

摘　　要：目的:制备一种以sLAG-3为佐剂的过敏原疫苗,并对该蛋白进行生物学活性测定,为建立一种高效安全的过敏原特异性免疫治疗奠定基础。方法:将sLAG-3和Fc段基因片段克隆入pcDNA3.1真核表达载体,电穿孔转染细胞,Western blotting检测生物活性。结果:成功构建可高度表达sLAG-3-Ig的真核表达载体,该蛋白可以明显抑制B细胞合成IgE的能力。结论:制备了一种以sLAG-3为佐剂的过敏原疫苗,该蛋白具有淋巴细胞抑制活性,从而可能建立一种高效安全的过敏原特异性免疫治疗手段。AIM： To obtain a vaccine with sLAG - 3 as immunoadjuvant and investigate its biologic activity in order to establish the safe and effective way for asthma as one of the specific immunotherapy. METHODS： The coding sequence of LAG- 3 was amplified by polymerase chain reaction, the expression vector pcDNA- sLAG- 3 - Ig was constructed by inserting the PCR products of sLAG- 3 and Fe sequence of IgG. With electroporation transfection, pcDNA - sLAG - 3 - Ig was transfected into COS - 7 cells and its biologic activity was investigated by Western blotting analysis, RESULTS： By temperature induction, the LAG - 3 - Ig was highly expressed in E. coli DH5α, LAG- 3 - Ig fusion protein was observed by SDS- PAGE and Western blotting, the results showed that the LAG - 3 - Ig protein was an antagonist of the IL- 4 - induced synthesis of IgE in B cells. CONCLUSION： A new vaccine with sLAG - 3 as immunoadjuvant was obtained. It could inhibit synthesis of IgE in B cells. Thus, LAG - 3 - Ig would be hopeful to establish the safe and effective way for asthma as one of the specific immunotherapy.

关 键 词：sLAG-3 Fc段 疫苗 哮喘 

分 类 号：R363[医药卫生—病理学]