Autoimmunity to Bile Duct Epithelial Cells in Patients with Primary Billary Cirrhosis

原发性胆汁性肝硬变(PBC)是一种以进行性和炎症性肝内胆管闭塞为特征的神秘的慢性肝病，自身免疫介导的发病机制被认为是靶组织损伤的原因。我们报道了T细胞免疫在PBC发病机制中的主导作用，因为在白细胞迁移抑制试验中，外周血淋巴细胞对部分纯化的胆汁抗原致敏，而在PBC患者的血清中未检测到针对同一抗原的抗体。此外，在10小时的效靶比分别为100和200时，脾T淋巴细胞对自体胆管上皮(BE)细胞有细胞毒作用，而非T淋巴细胞则无此作用。采用…技术从尸检标本中获得BE细胞更多地泵入PBS液，辅以EDTA、DNAase和Disparse进入胆管。细胞毒性的特异性通过使用自体BE细胞的冷靶向抑制来显示。用OKT8或OKT4单抗进行补体裂解的阴性选择显示，效应器表型为CD8阳性。然后，用兔抗人胆管上皮细胞特异性抗原检测部分纯化的胆汁抗原，通过SDS-PAGE和Western blotting鉴定胆管上皮抗原。检测到相对分子质量大于200kD的B1、B2和B3抗原。将它们分别从凝胶上电洗脱下来，作为抗原刺激T细胞增殖。在1.0~0.1mCg/ml的浓度范围内，仅在PBC中检测到向B3的转化，而在其他肝病中未检测到，提示T细胞免疫在PBC的发病机制中起重要作用。较少

Primary biliary cirrhosis (PBC) is an enigmatic chronic liver disease characterized by progressive and inflammatory obliteration of intrahepatic bile ducts where autoimmune-mediated pathogenesis is considered to be responsible for damages to the target tissue. We reported a predominant role of T cell mediated immunity in the pathogenesis of PBC, because peripheral lymphocytes were sensitized to partially purified biliary antigen in leukocyte migration inhibition test, while antibody to the same antigen was not detected in the sera of PBC patients.In this study, histoimmunochemical investigation showed that OKT8 positive and Leul5 negative T cell population, phenotypically cytotoxic T cells infiltrated into damaged bile ducts in PBC. Furthermore, splenic T lymphocytes but not non-T lymphocytes were shown to be cytotoxic against autologous biliary epithelial( BE ) cells at effector to target ratios of 100 and 200 in 10hrs'^<51>Cr% release. BE cells were obtained from autopsy specimen by … More pumping PBS solution supplemented with EDTA, DNAase and Disparse into bile ducts. Specificity of cytotoxicity was shown by cold target inhibition using autologous BE cells. A phenotype of effectors was shown to be CD8 positive by negative selection using complement lysis with OKT8 or OKT4 monoclonal antibody. Next, biliary epithelial antigens were identified by SDS-PAGE and Western blotting of partially purified biliary antigen in which rabbit anti-biliary antigen specific to human biliary epithelial cells was used to detect the antigens. B1, B2 and B3 antigens with more than 200 KD mw were detected. They were eluted electophoretically from gels separately and used as antigens to stimulate T cell proliferation. Blastgensis to B1 at a constriction of 1.0 to 0.1 mc g/ml was detected exclusively in PBC but not in other liver diseases while blastgenesis to B3 was not detected in PBC.These data strongly point to the role of T cell immunity against biliary epithelial antigen; B1 antigen in the pathogenesis of PBC. Less