MCMV感染同种异型皮肤移植小鼠间质性肺炎模型的建立  

A model of interstitial pneumonitis by murine cytomegalovirusinfection in mice with allogenic skin transplantation

作　　者：倪德群[1] 赵纪强[2] 张玮 甘霖[1] 陈敬贤[1,3] 王明丽[1] 

机构地区：[1]安徽医科大学基础医学院微生物学教研室,合肥230032 [2]中山大学第一附属医院肾移植科,广州510080 [3]哥伦比亚大学病理与细胞生物学教研室,纽约10032

出　　处：《安徽医科大学学报》2011年第6期509-514,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:30872253);安徽省科技攻关项目(编号:08010302179)

摘　　要：目的建立小鼠巨细胞病毒(MCMV)经鼻腔急性感染同种异型皮肤移植BALB/c小鼠间质性肺炎模型。方法①25只供体C57BL/6雌鼠与100只受体BALB/c雌鼠背部皮肤移植后,每只小鼠腹腔注射环孢素A(CsA,12 mg/kg),连续14 d。②移植受体小鼠随机分为5组,其中4组每只鼻腔接种30μl不同剂量MCMV悬液(102、103、104、105PFU/只),另一组为正常对照组,小鼠鼻腔接种原代小鼠胚胎成纤维细胞(MEF)悬液30μl/只。在接种后第5、9、14、21天处死小鼠,无菌取肺组织分别做HE染色、透射电镜、PCR、RT-PCR检测MCMV Smith毒株的即刻早期(IE)和晚期基因糖蛋白B(gB)基因转录产物,并进行原位杂交和免疫组织化学实验。结果 104、105 PFU实验组小鼠肺组织有局灶性的病理损害,并发现感染后第14天肺组织病理改变最明显;透射电镜检测到疱疹样病毒颗粒;PCR、RT-PCR检测实验组MCMV IE和gB基因及其转录产物均为阳性;两组原位杂交和免疫组织化学检测均可见肺间质上皮细胞中存在病毒核酸和蛋白;并在感染后的21 d内,105 PFU组小鼠相比对照组有明显的临床表现。结论成功建立同种异型皮肤移植后MCMV急性感染所致的小鼠间质性肺炎模型。Objective To establish a mouse model of interstitial pneumonitis with acute murine eytomegalovirus（MCMV） after allogeneic skin transplantation. Methods ① 25 of C57BL/6 donor mice and 100 of BALB/c recipient mice were underwent back skin graft. Cyelosporine A （ CsA, 12 mg/kg） was subsequently given for 14 days. ② All recipient animals were randomly divided into five groups （n = 20）, four experimental groups were infected intranasally with MCMV infected mouse embryonic fibroblasts （MEF） at different concentrations （ 102, 103, 104 and 105 PFU） in a volume of 30 μl, and the control group was inoculated with 30 μ1 of MEF cell suspension. After infection days 5,9,14 and 21, mice were sacrificed and lung samples were collected for HE staining, transmission electron microscope （ TEM）, PCR/RT-PCR, in situ hybridization, and immunohistochemistry. Results Focal pathological abnormality in the lung was observed in the mice with high dose of virus （ 10^4 and 10^5 PFU）. Herpes virus particles were found by TEM in the epithelial of the lung tissue. MCMV DNA was detected by PCR for immediate early（ IE ） genes and late genes glycoprotein B （gB）, and it was localized in lung ceils by in situ hybrid- ization. Viral mRNA for IE and gB were also present suggesting lytic infection in the lung tissue. MCMV protein was revealed in the interstitial lung epithelial ceils by immunohistochemistry. Clinical manifestations were only ob-served in the group with 10^5 PFU in 21 days after infection. Conclusion The study demonstrates that a mouse model of interstitial pneumonitis is successfully established after allozeneic mouse skin transnlantation.

关 键 词：鼠巨细胞病毒属 疾病模型 动物 

分 类 号：R563.19[医药卫生—呼吸系统] R373.1[医药卫生—内科学]