Leung Patrick S C, Park Ogyi, Matsumura Shuji, Ansari Aftab A, Coppel Ross L, Gershwin M Eric
Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis, CA 95616, USA.
Clin Dev Immunol. 2003 Jun-Dec;10(2-4):227-33. doi: 10.1080/10446670310001642429.
Over the past two decades, a number of studies have failed to provide direct evidence of specific microbial chronic infection in primary biliary cirrhosis (PBC). However, a recent report suggests that there is a specific association of Chlamydia pneumoniae in patients with PBC and that C. pneumoniae or similar antigens might play a role in the pathogenesis of disease. To determine if Chlamydia infection is associated with PBC, we applied a combination of immunological and molecular approaches to investigate (a) the serological reactivity against two common Chlamydia human pathogens, C. pneumoniae and C. trachomatis, by immunoblotting, (b) the presence of Chlamydia in liver samples of patients with PBC and controls by PCR amplification of Chlamydia specific 16S rRNA and (c) the presence of Chlamydia proteins in liver samples of patients with PBC and controls by immunohistochemical staining. By immunoblotting, C. trachomatis and C. pneumoniae specific serological antibodies were found in 52/57 (91.2%) AMA positive PBC, 7/33 (21/2%) of AMA negative PBC, 1/25 (4%) PSC, 0/15 (0%) Sjorgen's syndrome and 0/20 (0%) systemic lupus erythematosus patients and 0/20 (0%) healthy volunteers at 1:200 sera dilution. PBC sera reacted to Chlamydia and E. coli lysates in western blots up to a maximum of 10(-4) dilution. However, PCR amplification of the Chlamydia specific 16S rRNA gene was negative in 25/25 PBC livers but positive in 1/4 PSC liver, 3/6 in other liver disease controls and 1/4 normal liver samples. While two commercially available specific monoclonal antibodies stained positive controls (Chlamydia infected HEp-2 cells) they failed to detect Chlamydia antigens in PBC livers. The detection of Chlamydia specific antibodies but not Chlamydia rRNA gene and Chlamydia antigens in PBC suggests that Chlamydia infection is not involved in PBC.
在过去二十年中，多项研究未能提供原发性胆汁性肝硬化（PBC）中特定微生物慢性感染的直接证据。然而，最近一份报告表明，PBC患者中肺炎衣原体存在特定关联，且肺炎衣原体或类似抗原可能在疾病发病机制中起作用。为确定衣原体感染是否与PBC相关，我们应用免疫和分子方法相结合来研究：（a）通过免疫印迹法检测针对两种常见的人衣原体病原体——肺炎衣原体和沙眼衣原体的血清学反应性；（b）通过对衣原体特异性16S rRNA进行PCR扩增，检测PBC患者和对照肝脏样本中衣原体的存在情况；（c）通过免疫组织化学染色，检测PBC患者和对照肝脏样本中衣原体蛋白的存在情况。通过免疫印迹法，在血清稀释度为1:200时，52/57（91.2%）抗线粒体抗体（AMA）阳性的PBC患者、7/33（21.2%）AMA阴性的PBC患者、1/25（4%）原发性硬化性胆管炎（PSC）患者、0/15（0%）干燥综合征患者、0/2（0%）系统性红斑狼疮患者以及0/20（0%）健康志愿者中检测到沙眼衣原体和肺炎衣原体特异性血清抗体。PBC血清在蛋白质免疫印迹中与衣原体和大肠杆菌裂解物反应，最高稀释度可达10^(-4)。然而，25/25例PBC肝脏样本中衣原体特异性16S rRNA基因的PCR扩增结果为阴性，而1/4例PSC肝脏样本、3/6例其他肝病对照样本以及1/4例正常肝脏样本的检测结果为阳性。虽然两种市售的特异性单克隆抗体对阳性对照（衣原体感染的人喉表皮样癌细胞）染色呈阳性，但它们未能在PBC肝脏中检测到衣原体抗原。PBC中检测到衣原体特异性抗体，但未检测到衣原体rRNA基因和衣原体抗原，这表明衣原体感染与PBC无关。
