可诱导肺部特异表达白介素22转基因小鼠模型的建立

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 周立 ZHOU Li 1 2 first-author 袁宝军 YUAN Baojun 2 周宇麒 ZHOU Yuqi 2 董佳佳 DONG Jiajia 2 3 方萍 FANG Ping 2 郑涛 ZHENG Tao 2 3 朱舟 ZHU Zhou 2 3 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 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 Animal Bio-Safety LevelⅢ Laboratory at the Center for Animal Experiment/State Key Laboratory of Virology,Wuhan University Animal Bio-Safety LevelⅢ Laboratory at the Center for Animal Experiment/State Key Laboratory of Virology,Wuhan University 约翰霍普金斯大学医学院过敏和临床免疫学系 约翰霍普金斯大学医学院过敏和临床免疫学系 Division of Allergy and Clinical Immunology,School of Medicine,Johns Hopkins University Division of Allergy and Clinical Immunology,School of Medicine,Johns Hopkins University 耶鲁大学医学院过敏和临床免疫学系 耶鲁大学医学院过敏和临床免疫学系 Section of Allergy and Clinical Immunology,School of Medicine,Yale University Section of Allergy and Clinical Immunology,School of Medicine,Yale University 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（ 4 00ng·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（ 4 00ng·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. 白介素22 强力霉素 Tet-on/TRE-Tight诱导表达系统 interleukin-22（IL-22） doxycycline Tet-on/TRE-Tight system 10.14188/j.1671-8836.2015.03.002 R-332 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 2015-03-01 2021-03-31 3 61