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目的 建立一个在大鼠关节滑膜细胞表达人白细胞介素１０（ｈｕｍａｎ ｉｎｔｅｒｌｅｕｋｉｎ １０ ，ｈ Ｉ Ｌ１０） 的重组逆转录病毒载体基因转移系统，为下一步的研究工作做准备。方法 构建表达人白细胞介素１０ 的逆转录病毒重组体ｐ Ｌ Ｘ（ｈ Ｉ Ｌ１０） Ｓ Ｎ，经 Ｐ Ａ３１７ 细胞包装， Ｇ４１８ 筛选， Ｎ Ｉ Ｈ３ Ｔ３ 细胞进行病毒滴度测定，选滴度最高的克隆（６ ×１０８ 集落形成单位／ Ｌ） 作为感染大鼠关节滑膜细胞的感染细胞；用重组逆转录病毒感染大鼠关节滑膜细胞，应用聚合酶链反应（ Ｐ Ｃ Ｒ） ，反转录聚合酶链反应（ Ｒ Ｔ Ｐ Ｃ Ｒ） 和 Ｅ Ｌ Ｉ Ｓ Ａ 检测ｈ Ｉ Ｌ１０ 基因的整合和表达。结果 外源性ｈ Ｉ Ｌ１０ 基因已整合到靶细胞染色体 Ｄ Ｎ Ａ 并有效地表达。 Ｅ Ｌ Ｉ Ｓ Ａ 检测显示ｈ Ｉ Ｌ１０ 基因在ｐ Ｌ Ｘ（ｈ Ｉ Ｌ１０） Ｓ Ｎ 转染大鼠关节滑膜细胞４８ 小时后开始表达，达７２０ ｎｇ·１０ － ６ｃｅｌｌｓ·２４ｈ － １ ，高峰在第３ 天，为１ ９８２ ｎｇ·１０ － ６ｃｅｌｌｓ·２４ｈ － １ 。第７ 、１４ 、２８天分别为１２ ７６１ 、１ ０５４ 、９４２ ｎｇ·１０ － ６ｃｅｌｌｓ·２４ｈ － １ 。结论 外源性ｈ Ｉ ?
Objective To establish a recombinant retroviral vector gene transfer system expressing human interleukin 10 (hIL-10) in synovial cells of rats, and to prepare for the further research work. Methods The retrovirus recombinant pL X (h I L10) S N expressing human interleukin 10 was constructed and packaged in P A317 cells, screened with G418, and N I H3 T3 cells were used for virus titer determination. The highest clone (6 × 108 colony forming units / L) was used as the infected cells in the synovial cells of infected rats. The rat synovial cells were infected with recombinant retrovirus. PCR (P C R) Transcriptional Polymerase Chain Reaction (RT-P C R) and E L I S A Detection h I L 10 gene integration and expression. Results exogenous h I L 10 gene has been integrated into the target cell chromosome D N A and effectively expressed. E L I S A test showed that h I L10 gene began to express in synoviocytes of rat synovial cells after p L X (h I L10) S N transfection for 48 hours, reaching 720 ng · 10 -6 clls · 24h - 1, the peak on the third day was 1 982 ng · 10 - 6 cells · 24h - 1. The days 7, 14 and 28 were 12 761, 1054, 942 ng · 10 - 6 cells · 24h - 1, respectively. Conclusion Exogenous h I?