CAJ | 학술논문

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5.5×105 PFU的HCMV AD169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、Western blot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UL55 DNA；Western blot、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HCMV AD169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。
목적건립인거세포병독( HCMV)급성간질성폐염적소서모형。방법용5.5×105 PFU적HCMV AD169주미정맥주사6~8주SPF급BALB/c소서작위실험조，동시설립정상세포작위대조조，분별우접충후제5천화제30천처사소서，무균취소서폐조직，통과병독분리、PCR검측병독특정보수기인、면역조화、Western blot화HE염색적방법，관찰검측소서폐조직중HCMV급기조직병리변화。결과재감염후제5천화제30천실험조소서적폐조직중분리출료HC-MV병독；PCR검측도HCMV 특이성조만기기인IE화UL55 DNA；Western blot、면역조화검측도HCMV특이성조만기단백IE화gB；HE염색증실폐조직유명현급성간질성폐염적병리개변；대조조대응결과전위음성。결론통과미정맥주사HCMV AD169주병독，재감염후제5천성공구건료HCMV급성간질성폐염소서모형，해모형적건립유리우해충질병적발병궤제연구급상관항병독역묘화약물적연발。
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 5.5í105 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 gB 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.