抗酸性同功铁蛋白重组免疫细胞因子的制备及其特性研究  

Preparation and characterization of anti-acidic isoferritin recombinant immunocytokine

作　　者：游上游[1] 郭军庆[1] 艾洪武[1] 陈柳[1] 

机构地区：[1]武汉市儿童医院检验部,430016

出　　处：《中华微生物学和免疫学杂志》2005年第8期625-629,共5页Chinese Journal of Microbiology and Immunology

基　　金：武汉市科技局重点攻关计划(996009217)

摘　　要：目的构建并表达抗酸性同功铁蛋白(acidicisoferritin,AIF)的免疫细胞因子,并研究其抗肿瘤特性。方法在克隆小鼠趋化因子CXCL10全长基因的基础上,构建抗AIF单链抗体与CXCL10组成的重组免疫细胞因子,并在小鼠骨髓瘤细胞NSO中进行表达;使用流式细胞技术(FCM)和体外细胞趋化试验等方法来研究该重组蛋白的抗肿瘤特性。结果重组免疫细胞因子(IP10scFv)真核表达产物的相对分子质量(Mr)约为41.1×103,纯化后的蛋白浓度为68mgL,抗体亲和常数(KDIP10scFv)为2.28×10-8molL。IP10scFv能够特异识别分泌AIF的SMMC7721细胞,并能与激活后的小鼠T淋巴细胞表面CXCR3受体特异性结合,对小鼠激活的T淋巴细胞有较强的趋化作用。结论成功地制备了抗AIF单链抗体与趋化因子CXCL10构成的重组免疫细胞因子,该免疫细胞因子是肿瘤治疗的潜在制剂。Objective To study anti-tumor activity of the immunocytokine consisting of anti-AIF singlechain Fv （scFv） and chemokine IP10. Methods Recombinant immunocytokine of anti-AIF and CXCL10 was constructed and expressed in NSO cell hne by cloning of mouse chemokine CXCL10 gene. Anti-tumor component of the immunocytokine, called IP10-scFv, was studied with flow cytometry, immuno-fluorescence assay and chemotaxis assay. Results The molecular relative mass（Mr） of immunocytokine, IP10-scFv is 41.1 kD, affinity constant of IP10-scFv is 2.28×10^-8 with Scatchard formula. Indirect immuno-fluorescence and flow cytometry assay proved that IP10-scFv recognized both of SMMC 7721 cell hne secreting AIF and murine activated T cell that expressing CXCR3. Chemotaxis assay showed that IP10-scFv was a strong chemotaxis component from activated lymphocytes in vitro. Conclusion Immunocytokine IP10-scFv is a potential anti-tumor bioproduct.

关 键 词：酸性同功铁蛋白 免疫细胞因子 肿瘤 重组蛋白 特性研究 小鼠骨髓瘤细胞 制备 SMMC7721细胞 抗肿瘤特性 T淋巴细胞 

分 类 号：R392[医药卫生—免疫学]