HCMV急性间质性肺炎小鼠模型的建立  

A murine model of acute interstitial pneumonia induced by primary infection of human cytomegalovirus

作　　者：庄晓亮[1] 木朝宇 俞俊岭 张文昌[1] 甘霖[1] 陈敬贤[1,2] 王明丽[1] 

机构地区：[1]安徽医科大学微生物学教研室,合肥230032 [2]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》2014年第9期1214-1217,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:30872253);安徽省科技厅科技攻关项目(编号:8010302179);安徽省教育厅自然科学重点课题(编号:KJ2009A041Z)

摘　　要：目的建立人巨细胞病毒（HCMV）急性间质性肺炎的小鼠模型。方法用5．5×105PFU的HCMVADl69株尾静脉注射6～8周SPF级BALB／c小鼠作为实验组，同时设立正常细胞作为对照组，分别于接种后第5天和第30天处死小鼠，无菌取小鼠肺组织，通过病毒分离、PCR检测病毒特定保守基因、免疫组化、Westernblot和HE染色的方法，观察检测小鼠肺组织中HCMV及其组织病理变化。结果在感染后第5天和第30天实验组小鼠的肺组织中分离出了HC—MV病毒；PCR检测到HCMV特异性早晚期基因IE和UL55DNA；Westernblot、免疫组化检测到HCMV特异性早晚期蛋白IE和gB；HE染色证实肺组织有明显急性间质性肺炎的病理改变；对照组对应结果全为阴性。结论通过尾静脉注射HCMVADl69株病毒，在感染后第5天成功构建了HCMV急性间质性肺炎小鼠模型，该模型的建立有利于该种疾病的发病机制研究及相关抗病毒疫苗和药物的研发。Objective To establish the model of acute interstitial pneumonia induced by primary infection of human cytomegalovirus （ HCMV）, and to provide a reliable platform for the research on pathogenesis of HCMV disease and for the estimate of antivirals and vaccines. Methods Twenty-four specific-pathogen-free BALB/c mice of 6 ～ 8 weeks old were randomly divided into two groups. The infection groups were injected with HCMV AD169-infected HF at 5.5 x 105 PFU by intravenous injection respectively, and the control group was injected with HF only. On the post infection days 5 and 30, mice were anesthesia executed and lung samples were collected for analysis by vi- ral isolation, polymerase chain reaction （ PCR）, immunohistochemistry, Western blot and HE staining, to observe the correlation between HCMV and pathological change in lung of mice. Results After infection 5 d and 30 d, HCMV was only isolated from the lung homogenates of the infection group, the HCMV IE-1 and UL55 DNA were positive by PCR, and the viral major proteins, IE and antigen, were widely distributed in the interstitial lung epithelial cells by immunohistochemistry and revealed in the lung by Western blot in the group of infection, which were negative in the control group. The obvious pathological changes of acute interstitial pneumonia were found in the lung tissue of the infection group by HE staining. Conclusion The murine model of acute interstitial pneumo- nia is successfully established which only costs 5 days after intravenous injection of HCMV, providing an important platform for pathogenesis of HCMV acute interstitial pneumonitis and medicine intervention.

关 键 词：人巨细胞病毒 小鼠模型 间质性肺炎 

分 类 号：R563.13[医药卫生—呼吸系统] R373.9[医药卫生—内科学]