人OX40转基因细胞株的建立及其对树突状细胞的促分化作用  

Construction of Human OX40 Transfectant and Its Potency to Induce Differentiation of Dendritic Cells

作　　者：孙建军[1] 施勤[1] 王勤[1] 朱一蓓[1] 陈洁[1] 陈永井[1] 张学光[1] 

机构地区：[1]苏州大学生物技术研究所,苏州215007

出　　处：《上海免疫学杂志》2003年第6期370-373,共4页Shanghai Journal of Immunology

基　　金：国家重大基础研究资助项目 (No 2 0 0 1CB5 10 0 0 3 ) ;江苏省自然基金重点资助项目 (No BJ9810 0 )

摘　　要：应用RT PCR方法 ,从PHA活化的扁桃体T细胞的mRNA中 ,扩增获得编码人OX4 0分子的cDNA ,并将其克隆到pMD18 T载体 ,经PCR、酶切和测序鉴定确证。进而构建pcDNA3 1 OX4 0重组真核表达载体 ,脂质体法转染L92 9细胞 ,经G4 18筛选 ,获得能稳定高表达OX4 0分子的细胞株。经OX4 0 FITC单抗标记和FCM检测 ,阳性表达率为 89 1%。转基因细胞在可溶性CD4 0L共存条件下 ,与未成熟DC共育 ,能促进DC的进一步分化成熟 ,使DC膜分子CD4 0、CD86和CD83呈上调性表达。The cDNA fragment encoding the human OX40 was obtained from the mRNA of PHA-activated T cells of tonsil by using RT-PCR and cloned into the pMD18-T vector. By DNA sequencing,the cDNA was consistent with the reported human OX40 cDNA in the GenBank,and then was inserted into the eukaryotic expression vector pcDNA3.1. The recombinant plasmid was transfected into L929 cell line with lipofectine,selected by G418 after transfection,a stable cell line expressing the human OX40 was established. The expression stability and efficiency of the target molecule was assayed by immunophenotyping and flow cytometry analysis. It was found that the positive rate of expression was 89.1%. In the presence of sCD40L,OX40 transfectant was evidenced to have a potency to induce the differentiation of dendritic cells in vitro and to up-regulate the expression of maturation markers,such as CD40,CD86 and CD83.

关 键 词：人OX40 基因转染 共刺激分子 树突状细胞 

分 类 号：R392.11[医药卫生—免疫学]