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  

Analysis on Promoter Activities of IL-10,IL-4 and their receptors in replication of KSHV induced by HSV-1

作　　者：吕志刚[1] 卢春[1] 秦娣[1] 曾怡[1] 程林[1] 陈秀英[1] 

机构地区：[1]南京医科大学微生物学与免疫学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2008年第2期139-143,193,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：霍英东青年教师基金资助项目(101038)

摘　　要：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。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-4Rα 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-l-induced KSHV replication in BCBL-1 cells.

关 键 词：卡波济肉瘤相关疱疹病毒 人类单纯疱疹病毒1型 白细胞介素 启动子活性 

分 类 号：R373[医药卫生—病原生物学]