可诱导肺部特异表达白介素22转基因小鼠模型的建立  

Establishment of a Transgenic Mouse Model with Lung-Specific Expression of IL-22 Using the Tet-on Inducible System

作　　者：周立[1,2] 袁宝军[2] 周宇麒[2] 董佳佳[2,3] 方萍[2] 郑涛[2,3] 朱舟[2,3] 

机构地区：[1]武汉大学病毒学国家重点实验室动物实验中心/ABSL-Ⅲ实验室,湖北武汉430071 [2]约翰霍普金斯大学医学院过敏和临床免疫学系,马里兰巴尔的摩21224 [3]耶鲁大学医学院过敏和临床免疫学系,康涅狄格纽黑文06511

出　　处：《武汉大学学报（理学版）》2015年第3期203-207,共5页Journal of Wuhan University:Natural Science Edition

基　　金：中国博士后科学基金面上项目(2013M531745);国家自然科学基金(81301428);中央高校基本科研业务费专项资金(2042015kf0188);美国国立卫生研究院项目(R01HL079349;P01HL107151;R01AI075025)资助

摘　　要：建立了一种肺部特异性、高效表达鼠源白介素22（mIL-22）的转基因小鼠模型.将TRE-Tight-mIL-22转基因片段显微注射入C57BL/6J小鼠受精卵细胞中,得到转基因小鼠,与能够控制TRE-Tight下游基因在肺部特异性表达的CC10-rtTA-hGH转基因小鼠交配,获得同时含有CC10-rtTA-hGH和TRE-Tight-mIL-22这两段基因的双阳性子代转基因小鼠（IL-22转基因小鼠）.使用强力霉素（Dox）持续诱导6周龄IL-22转基因小鼠6周,用酶联免疫吸附测定法（ELISA）检测小鼠肺部灌洗液中IL-22蛋白的表达量.结果表明,在野生型和未诱导的IL-22转基因小鼠的肺部均未发现IL-22表达,而经Dox诱导后,与野生型C57BL/6J小鼠（〈10ng·L-1）比较,IL-22转基因小鼠肺部发现高表达量IL-22（~400ng·L-1,p〈0.05）.免疫组化染色结果显示,经Dox诱导,IL-22主要在IL-22转基因小鼠的肺部气道上皮细胞特异性表达.An inducible lung-specific Interleukin-22（IL-22）expressing transgenic mouse model with high level expression of IL-22 in the lung was established.Transgenic mice were generated by pro-nuclear microinjection of the transgenic construct TRE-Tight-mIL-22,and crossbreeding TRE-Tight-mIL-22 positive mice with CC10-rtTA-hGH transgenic mice,which contain the promoter that could control the expression of gene of interest specifically in the lung epithelial cells.Then the double positive transgenic（Tg）mice that contained both TRE-Tight-mIL-22 and CC10-rtTA-hGH genes were obtained.Starting at 6weeks of age,doxycycline（Dox）was given in the drinking water to turn on the IL-22 transgene for 6weeks,and the concentration of IL-22 protein in the bronchoalveolar lavage（BAL）fluid and serum samples of these Tg mice and wild type control mice was determined by enzyme-linked immunosorbent assay（ELISA）.Without Dox,no IL-22 was detected in the BAL fluid of wild type or IL-22 Tg mice.However,double positive IL-22 Tg mice given Dox had high level expression of IL-22 in the BAL samples compared with wild type mice（~400ng·L-1 vs.〈10ng·L-1,p〈0.05）.Immunohistochemistry（IHC）using anti-IL-22 antibody dem-onstrated that IL-22 was expressed in the airway epithelial cells of TRE-Tight-mIL-22 Tg mice after Dox induction only.

关 键 词：白介素22 强力霉素 Tet-on/TRE-Tight诱导表达系统 

分 类 号：R392-3[医药卫生—免疫学]