Marangoni Antonella, Donati Manuela, Cavrini Francesca, Aldini Rita, Accardo Silvia, Sambri Vittorio, Montagnani Marco, Cevenini Roberto
Sezione di Microbiologia DMCSS, University of Bologna, Policlinico S.Orsola, 9 Via Massarenti, Bologna 40138, Italy.
World J Gastroenterol. 2006 Oct 28;12(40):6453-7. doi: 10.3748/wjg.v12.i40.6453.
To develop an animal model of liver infection with Chlamydia pneumoniae (C. pneumoniae) in intraperitoneally infected mice for studying the presence of chlamydiae in Kupffer cells and hepatocytes.
A total of 80 BALB/c mice were inoculated intraperitoneally with C. pneumoniae and sacrificed at various time points after infection. Chlamydiae were looked for in liver homogenates as well as in Kupffer cells and hepatocytes separated by liver perfusion with collagenase. C. pneumoniae was detected by both isolation in LLC-MK2 cells and fluorescence in situ hybridization (FISH). The releasing of TNFA-alpha by C. pneumoniae in vitro stimulated Kupffer cells was studied by enzyme-linked immunosorbent assay.
C. pneumoniae isolation from liver homogenates reached a plateau on d 7 after infection when 6 of 10 animals were positive, then decreased, and became negative by d 20. C. pneumoniae isolation from separated Kupffer cells reached a plateau on d 7 when 5 of 10 animals were positive, and became negative by d 20. The detection of C. pneumoniae in separated Kupffer cells by FISH, confirmed the results obtained by culture. Isolated hepatocytes were always negative. Stimulation of Kupffer cells by alive C. pneumoniae elicited high TNF-alpha levels.
A productive infection by C. pneumoniae may take place in Kupffer cells and C. pneumoniae induces a local pro-inflammatory activity. C. pneumoniae is therefore, able to act as antigenic stimulus when localized in the liver. One could speculate that C. pneumoniae infection, involving cells of the innate immunity such as Kupffer cells, could also trigger pathological immune reactions involving the liver, as observed in human patients with primary biliary cirrhosis.
通过腹腔感染小鼠建立肺炎衣原体（C. pneumoniae）肝脏感染动物模型，以研究库普弗细胞和肝细胞中衣原体的存在情况。
将80只BALB/c小鼠腹腔接种肺炎衣原体，并在感染后的不同时间点处死。在肝脏匀浆以及通过胶原酶灌注肝脏分离得到的库普弗细胞和肝细胞中寻找衣原体。通过在LLC-MK2细胞中分离培养以及荧光原位杂交（FISH）检测肺炎衣原体。采用酶联免疫吸附测定法研究体外受肺炎衣原体刺激的库普弗细胞中肿瘤坏死因子-α（TNFA-α）的释放情况。
感染后第7天，从肝脏匀浆中分离出肺炎衣原体的阳性率达到平台期，10只动物中有6只呈阳性，随后下降，到第20天时转阴。从分离得到的库普弗细胞中分离出肺炎衣原体的阳性率在第7天达到平台期，10只动物中有5只呈阳性，到第20天时转阴。通过FISH检测分离得到的库普弗细胞中的肺炎衣原体，证实了培养结果。分离得到的肝细胞始终为阴性。活的肺炎衣原体刺激库普弗细胞可引发较高的肿瘤坏死因子-α水平。
肺炎衣原体可在库普弗细胞中发生有效感染，且肺炎衣原体可诱导局部促炎活性。因此，肺炎衣原体定位于肝脏时能够作为抗原刺激物。可以推测，如在原发性胆汁性肝硬化人类患者中所观察到的那样，涉及库普弗细胞等天然免疫细胞的肺炎衣原体感染也可能引发涉及肝脏的病理性免疫反应。
