一种新型炎症诱导型腺病毒表达系统的功能研究  

A functional analysis for a new inflammation-inducible adenoviral expression system in vitro and in vivo

作　　者：蔡刚[1] 聂小蒙[2] 管政[1] 张军[1] 沈茜[1] 

机构地区：[1]第二军医大学附属长海医院实验诊断科,上海200433 [2]第二军医大学附属长海医院呼吸内科

出　　处：《中华微生物学和免疫学杂志》2007年第4期337-342,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助（No.39970270）

摘　　要：目的 构建一种新型炎症诱导型重组腺病毒载体用于生理调控型基因治疗。方法 构建携带IL-1β增强子（-3690／-2720）／IL-6启动子（-590／＋2）重组体（IL1／IL6）的荧光素酶报告质粒和重组腺病毒载体，体外转染HepG2和RAW264．7细胞，观察各种细胞外刺激对IL1／IL6启动子活性调控的变化，并以动物实验观察炎症因子能否在体内通过调控IL1／IL6的启动活性而控制目的基因的表达。结果经荧光素酶分析显示IL1／IL6的本底启动活性较低，可经多种细胞外刺激而诱导活化，且具有较广泛的活性诱导范围；在体内，它可随炎症反应程度的变化而自主地调控外源基因的表达。结论携带IL1／IL6重组体启动子的重组腺病毒载体可作为生理反应调控型基因治疗载体用于全身性炎症、自身免疫性疾病等的治疗与研究。Objective To develop an inflammation-regulated recombinant adenoviral expression system for physiologically responsive gene therapy.Methods A luciferase reporter vector with a hybrid promoter containing human IL-1β enhancer region （-3690 to -2720） and human IL-6 promoter （-590 to ＋2） was constructed. A replication-deficient adenoviros was engineered with luciferase controlled by the IL1/IL6 promoter （Ad-IL1/IL6-Luc）. The reporter vector or adenovirns was transfected to HepG2 or RAW264.7 cells to study the in vitro charactors of this hybrid promoter. Inflammation model was prepared by intraperitoneally injecting BALB/c mice with LPS, and intravenously infecting them with Ad-IL1/IL6-Luc or Ad-CMV-Luc to study the in vivo characters of ILl/ IL6 promoter. Results The ILI/IL6 hybrid promoter has pronounced promoter activity, has broad-range responsiveness to extracellular signaling molecules,and can be re-challenged after first induction. The IL1/IL6 could drive the transgene expression parallelling with the disease progression-upregulation in relapsing and downregulation in restoration.Conclusion The IL1/IL6 promoter fidfills the criteria of a disease-inducible promoter： low basal activity and high during the acute responsive phase. This promoter might be feasible for auto-regulated transgene products treatment of autoimmune diseases and systemic inflammation diseases using adenoviral gene therapy.

关 键 词：基因治疗 腺病毒载体 可调控型启动子 炎症反应 

分 类 号：R686[医药卫生—骨科学]