小鼠的原发性呼吸道合胞病毒感染

Primary respiratory syncytial virus infection in mice

目的 研究呼吸道合胞病毒 (RSV)感染模型的建立。方法 实验组用在HEP 2细胞上培养出的不同滴度 (7 5× 10 5,2 75× 10 6,2 5× 10 7PFU/ml)的 9815 9株RSV予Balb/c小鼠滴鼻 ,每个滴度 3只小鼠 ,共 9只。对照组 3只小鼠用HEP 2细胞悬液滴鼻 ,5d后 ,无菌状态下取肺 ,右肺称重 ,按1∶10加入eagleMEM ,匀浆 ,4℃ ,2 0 0 0r/min离心 2 0min ,上清液加入HEP 2细胞作病毒分离 ,空斑分析。左肺用于病理检查 ,电镜检查。结果 (1)RSV感染使梭形的HEP 2细胞融合成地图状、圆形、椭圆形、树枝状 ;(2 )肺组织匀浆接种于HEP 2细胞上 ,于接种的 10～ 12d分离出病毒 ;(3)不同滴度的RSV在肺组织中的复制滴度不同 ,滴度越高 ,肺复制滴度越高 ;(4 )较高滴度的RSV感染小鼠 ,肺组织病毒复制于感染后的 4～ 6d达高峰 ,肺病理变化于 5～ 8d最明显 ,可见炎症细胞浸润 ,主要是淋巴细胞 ;低滴度RSV感染小鼠后病理变化不明显 ;(5 )电镜检查 :高倍镜下可见毛刺样外观的病毒 ,病变呈横切或纵切。结论 采用 9815

Objective To establish a mice-model of primary respiratory syncytial virus infection. Methods Twelve Balb/c mice (8-12 w) were divided into two g roups with three different titers of RSV infection in the experimental group M ice were anaesthetized with pentobabitone(30 mg/kg) and infected b y na sal drip of 01 ml of RSV which was harvested from HEP-2 cell culture Three mice in the control group received sham infection with HEP-2 cell media Five days later,lungs were removed in aseptic condition Right lung was weighted an d homoginized with MEM(1∶10 ratio),then centrifuged at 4℃,2 000 r/min for 20 minutesSupernatants were collected for virus isolation and plaque forming a ss ay Left lung was collected for histopathological and electromicroscopic exami nationResults (1) RSV grew wit h a characteristic syncytial fomation in HEP-2 cells, leading to the fusion of c ells with round, elliptic, map or tree like shape (2) Virus was isolated and dete cted in the lung tissue in experiment group(3) There was a positive correlatio n b etween RSV titer added and that replicated in the lung(4) After the eclipse ph ase , viral replication in the lung reached maximum within 4 and 6 days The most ap parent pathological change was found between day 5 and 8 Mononuclear cell infi l tration was presented in the perivascular, peribronchial and in the alveoli spa c ers(5) Huge viral replication with its fuzzy coat was detected inside the cyto p lasm of the type Ⅱ alveolar lining cell at high magnificationConcl usion An animal model of RSV infection Balb/c mice was successfully established by nasal inoculation with respiratory syncytial virus