IL-10?IL-4及其受体启动子在人类单纯疱疹病毒1型激活卡波济肉瘤相关疱疹病毒复制过程中的启动活性分析
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霍英东青年教师基金资助项目(101038)


Analysis on Promoter Activities of IL-10,IL-4 and their receptors in replication of KSHV induced by HSV-1
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    目的:分离和鉴定人类IL-10-IL-10受体(IL-10Rα)-IL-4及其受体IL-4Rα启动子序列,并分析IL-10-IL-4以及其受体的启动子在人类单纯疱疹病毒1型(HSV-1)激活卡波济肉瘤相关疱疹病毒(KSHV)过程中的启动活性-方法:以人类基因组DNA为模板,PCR扩增IL-10-IL-10Rα-IL-4Rα启动子区序列,并分别克隆载入pGL-3基本载体中虫荧光素酶(Luciferase)报告基因的上游-进一步将上述构建的重组质粒与本室保存的含IL-4启动子重组质粒分别转染HSV-1感染的BCBL-1细胞,并作Luciferase活性检测-结果:分离了IL-10-IL-10Rα和IL-4Rα的启动子区域序列,并成功克隆了含启动子序列重组报告质粒;HSV-1感染的BCBL-1细胞在进一步转染了IL-10-IL-10Rα-IL-4和IL-4Rα启动子重组报告质粒后,其Luciferase值与相应的对照比较显著升高(P < 0.05)-结论:在BCBL-1细胞中,HSV-1可通过直接激活IL-10-IL-4以及相应受体的启动子来上调它们的表达;在HSV-1感染的BCBL-1细胞中,IL-10和IL-4可能对KSHV的裂解复制起到促进作用-

    Abstract:

    Objective:To evaluate the promoters’ activities of human interleukin-10(IL-10),IL-10 receptor(IL-10Rα),IL-4 and IL-4 receptor(IL-4Rα) during the lytic cycle replication of Kaposi’s sarcoma-associated herpesvirus induced by herpes simplex virus type 1 (HSV-1) in BCBL-1 cells. Methods:Promoters’ sequences of IL-10,IL-10Rα and IL-4Ra were amplified using PCR from total DNA of human genome. The PCR products were further cloned into pGL-3 basic vector to construct recombinant luciferase reporter plasmids. After infected by HSV-1,BCBL-1 cells were transfected with the promoter-driven luciferase constructs to induce luciferase gene expression and calculate the relative luciferase activity unit(RLU). Results:IL-10,IL-10Rα and IL-4Rα promoter sequences were isolated and constructed recombinant luciferase reporter plasmids successfully. The luciferase reporter recombinant exhibited a strong promoter activity,and the promoter-driven luciferase expression was increased significantly after infected with HSV-1 in BCBL-1 cells when compared with pGL-3 basic vector(P < 0.05). Conclusion:The data presented the evidence that HSV-1 could up-regulate the expression of IL-10 and IL-4 by activating IL-10,IL-4 and their receptors. And these data also suggested that IL-10 and IL-4 should partially contribute to HSV-1-induced KSHV replication in BCBL-1 cells.

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吕志刚,卢 春,秦 娣,曾 怡,程 林,陈秀英. IL-10?IL-4及其受体启动子在人类单纯疱疹病毒1型激活卡波济肉瘤相关疱疹病毒复制过程中的启动活性分析[J].南京医科大学学报(自然科学版),2008,28(2):139-143193

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  • 收稿日期:2007-08-22
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