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.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
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)
