Zhang Wei, Gan Lin, Li Rui-ping, Chen Jing-xian, Wang Ming-li
Microbiology Department of Anhui Medical University, Hefei, 230032, China.
Bing Du Xue Bao. 2011 Jan;27(1):1-10.
To provide a reliable animal model for study of human CMV disease in gastrointestinal track, we tried to infect with murine cytomegalovirus (MCMV) in mice that were received allogenetic skin transplantation under immunosuppression. (1) Skin transplantation was performed between 18 donor C57BL/6 mice and 72 recipient BALB/c mice. (2) All recipient mice were then given Cyclosporine at 12 mg/kg daily for 2 weeks by intraperitoneal injection. Mice were randomly divided into 3 groups. Two experimental groups were received MCMV-infected mouse embryonic fibroblasts (MEF) at 10(4) PFU and 10(5) PFU respectively, and the control group received MEF only. We observed any possibly pathophysiological behavior changes and recorded the changes in body weight. The mice were sacrificed at 5d, 9d, 14d, 21d post infection and colon tissue was collected for analysis.
Mice infected with MCMV at 10(5) PFU group showed anorexia, lethargy and degression in locomotor activity. This group of mice showed significant decrease in body weight than that of other groups. Colon tissues were collected 14 days after infection. Histological examination revealed that the mucous layer became thinner in the proximal colon and increased number of lymphoid follicles in distal colon in infected animals. The changes in the mucosal structure was most prominent in the group 10(5) PFU MCMV. Viral DNA was present in the colon by in situ hybridization for IE1 gene, and viral gB transcript was positive by RT-PCR. One of the viral major proteins, pp65, was widely distributed in the colon by immunohistochemistry. These data demonstrated that MCMV established infection in colon of the mice after allogenetic skin transplantation. Electron microscopy showed that there were herpes virus particles in the colon tissue.
Infection with MCMV in mouse after allogenetic skin transplantation by nasal cavity inoculation resulted in the pathological changes in colon tissue similar to that of inflammation in human colon. The small animal model of colon inflammation may provide a platform for further study of pathogenesis as well as medical intervention of HCMV involved inflammation of human bowel.
为了提供一种可靠的动物模型用于研究人类巨细胞病毒在胃肠道的疾病,我们尝试在免疫抑制下接受同种异体皮肤移植的小鼠中感染鼠巨细胞病毒(MCMV)。(1)在18只供体C57BL/6小鼠和72只受体BALB/c小鼠之间进行皮肤移植。(2)然后所有受体小鼠通过腹腔注射每天给予12mg/kg环孢素,持续2周。小鼠被随机分为3组。两个实验组分别接受10⁴ PFU和10⁵ PFU的MCMV感染的小鼠胚胎成纤维细胞(MEF),对照组仅接受MEF。我们观察任何可能的病理生理行为变化并记录体重变化。在感染后5天、9天、14天、21天处死小鼠并收集结肠组织进行分析。
感染10⁵ PFU MCMV的小鼠组出现厌食、嗜睡和运动活动减退。该组小鼠体重比其他组显著下降。在感染后14天收集结肠组织。组织学检查显示,感染动物近端结肠的黏液层变薄,远端结肠的淋巴滤泡数量增加。黏膜结构变化在10⁵ PFU MCMV组最为显著。通过IE1基因原位杂交在结肠中检测到病毒DNA,通过RT-PCR检测到病毒gB转录本呈阳性。病毒主要蛋白之一pp65通过免疫组织化学在结肠中广泛分布。这些数据表明MCMV在同种异体皮肤移植后的小鼠结肠中建立了感染。电子显微镜显示结肠组织中有疱疹病毒颗粒。
通过鼻腔接种在同种异体皮肤移植后的小鼠中感染MCMV导致结肠组织出现类似于人类结肠炎症的病理变化。结肠炎症的小动物模型可能为进一步研究人巨细胞病毒相关肠道炎症的发病机制以及医学干预提供一个平台。