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| 鼠抗人CXCR3单克隆抗体的研制及生物学功能研究 | |
| 引用本文： | 黄莉,孙杰,郭静雅,邱玉华. 鼠抗人CXCR3单克隆抗体的研制及生物学功能研究[J]. 细胞与分子免疫学杂志, 2011, 27(12): 1325-1329 |
| 作者姓名： | 黄莉 孙杰 郭静雅 邱玉华 |
| 作者单位： | 1. 苏州大学附属儿童医院,江苏苏州,215003 2. 中国科学院苏州纳米技术与纳米仿生研究所,江苏苏州,215123 3. 苏州大学医学部免疫学系,江苏苏州,215123 |
| 基金项目： | 国家科技重大专项资助项目(2009ZX09103-705) |
| 摘 要： | 目的:获得持续分泌鼠抗人CXCR3单克隆抗体(mAb)的杂交瘤细胞株;以鼠抗人CXCR3 mAb作为工具,研究人CXCR3分子的表达特性及CXCR3信号转导对L929 -huCXCR3和结肠癌细胞株的迁移及增殖的影响.方法:以高表达人CXCR3膜型分子的L929-huCXCR3细胞作为免疫原免疫BALB/c小鼠,采用B淋巴细胞杂交技术,将免疫小鼠的脾脏细胞与同种系小鼠的骨髓瘤细胞sp2/0进行融合.以L929-huCXCR3作为筛选细胞,转染空载体的L929 -mock细胞作为阴性对照细胞,采用间接免疫荧光和流式细胞术,筛选能持续分泌抗人CXCR3 mAb的杂交瘤细胞株.采用Ig亚类快速定性试纸法和间接免疫荧光法对所获得的杂交瘤细胞株和mAb进行鉴定;用间接免疫荧光法分析CXCR3分子在肿瘤细胞表面的表达;Transwell隔离小室检测mAb对L929 -huCXCR3和结肠癌细胞株Colo205、HCT116及HT29迁移的影响;MTT法分析mAb对结肠癌细胞株Colo205增殖的影响.结果:获得了1株能持续分泌鼠抗人CXCR3 mAb的杂交瘤细胞株,命名为9B5.经快速定性试纸分析显示,该mAb重链为IgG1亚类,轻链为链;间接免疫荧光和流式细胞术分析显示,mAb 9B5可识别活化T淋巴细胞和结肠癌细胞株Colo205、HCT116及HT29表面的CXCR3分子.通过阻断CXCR3信号转导,mAb 9B5可抑制L929-huCXCR3细胞和结肠癌细胞株Colo205、HCT116和HT29的定向迁移及IP-10对Colo205的促增殖作用.结论:成功获得了1株能持续分泌鼠抗人CXCR3 mAb的杂交瘤细胞株,为研究CXCR3的表达特性及深入探讨CXCR3信号转导在肿瘤生长与转移过程中的作用及机制奠定了物质基础,并且有望为治疗肿瘤转移提供新的思路和新型药物. |
| 关 键 词： | CXCR3 单克隆抗体 趋化因子 肿瘤转移 细胞迁移 |
Mouse anti-human CXCR3 mAb preparation and its biological function |
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| HUANG Li , SUN Jie , GUO Jing-ya , QIU Yu-hua. Mouse anti-human CXCR3 mAb preparation and its biological function[J]. Chinese journal of cellular and molecular immunology, 2011, 27(12): 1325-1329 | |
| Authors: | HUANG Li SUN Jie GUO Jing-ya QIU Yu-hua |
| Affiliation: | HUANG Li1,SUN Jie2,GUO Jing-ya3,QIU Yu-hua3 1Children's Hospital Affiliated Soochow University,Suzhou 215003,2Division of Nano Biomedicine,Suzhou Institute of Nano-Tech and Nano-Bionics,Chinese Academy of Sciences,Suzhou 215123,3Department of Immunology,Medical College of Soochow University,China |
| Abstract: | AIM: Obtain hybridoma cell line with continuing expression of mouse anti-human CXCR3 mAb,investigate expression characteristics of human CXCR3 and how CXCR3 signal transduction function on L929-huCXCR3 and colon carcinoma cell lines transfer and growth.METHODS: Taking L929-huCXCR3 cell with high expression of human CXCR3 membrane molecule as immunogen to immunize BALB/c mouse,we fused immunized mouse spleen cell with myeloma cell Sp2/0 of its same germ line,then used L929-huCXCR3 as screening cell and empty... |
| Keywords: | CXCR3 monoclonal antibody chemokine tumor metastasis cell migration |
| 本文献已被 CNKI 万方数据 PubMed 等数据库收录！ | |