可溶性IL-6RB亚基突变体在CHO细胞中的表达及活性研究  

Expression of soluble interleukin-6 receptor B subunit mutant and studies of its activity

作　　者：杨志华[1] 倪长源[2] 沈倍奋[2] 王建安[2] 王之贤[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所 [2]军事医学科学院基础医学研究所，北京 100850

出　　处：《中华微生物学和免疫学杂志》2000年第6期580-583,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的　弄清只含白细胞介素 6 (IL 6 )受体 β亚基 (gp130 )胞外第二、三个FNⅢ区的可溶性突变体是否能拮抗IL 6的生物学效应。方法　首先用重叠延伸PCR方法扩增出gp130突变体cDNA ;接着将其克隆到真核表达载体pSVL中 ;用脂质体转染法将此表达载体导入CHO细胞中 ,通过点杂交和反转录PCR证明其得到有效表达 ;用MTT法在杂交瘤细胞和白血病细胞中观察了此突变体对IL 6生物学效应的影响。结果　发现此突变体的表达上清具有拮抗IL 6所致的杂交瘤细胞 7TD1增殖及白血病细胞R2生长抑制的作用。结论　gp130胞外第二、三个FNⅢ区中含有与IL 6 /IL 6Rα形成复合物的部位。此结果为深入研究gp130胞外区的结构与功能以及针对gp130胞外区的小分子激动剂和拮抗剂的设计提供了一些依据。Objective To investigate whether the soluble IL 6 receptor β chain (gp130) containing the second and the third fibronectin type Ⅲ modules of the extracellular region can antagonize the biological effects of IL 6. Methods The sgp130 mutant cDNA was amplified by using overlaping extension PCR, cloned into a mammalian expression vector, pSVL, and expressed in CHO cells. The effect of the expressed protein on IL 6 biological effects were studied in a hybridoma cell line (7TD1) and leukemic cell line (R2) systems. Results The sgp130 mutant can counteract the effects of IL 6 on the proliferation of 7TD1 and the growth inhibition of R2 cells. Conclusion The second and the third fibronectin type Ⅲ modules of the extracellular region may contain the binding sites of gp130 to IL 6 and IL 6Rα.

关 键 词：白细胞介素6 GP130 突变体 CHO细胞 表达 

分 类 号：R392.12[医药卫生—免疫学]