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

原发性胆汁性肝硬化（PBC）是一种神秘的慢性肝病，其特征是肝内胆管进行性和炎症性闭塞，其自身免疫介导的发病机制被认为是对靶组织损害的原因。我们报道了T细胞介导的免疫在PBC发病机制中的主要作用，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道抗原敏感，而在PBC患者的血清中未检测到针对同一抗原的抗体。在本研究中，组织免疫化学研究显示，OKT8阳性和Leul5阴性的T细胞群，表型上细胞毒性T细胞浸润到PBC损伤的胆管中。此外，脾T淋巴细胞而非非T淋巴细胞对自身胆道上皮（be）细胞具有细胞毒性，效应靶比为100和200,10hrs ^<51>Cr%释放。将添加EDTA、DNAase和Disparse的PBS溶液泵入胆管，从尸检标本中获得BE细胞。利用自体BE细胞进行冷靶抑制，显示了细胞毒性的特异性。通过与OKT8或OKT4单克隆抗体进行补体裂解的阴性选择，效应物的表型显示为CD8阳性。接下来，采用SDS-PAGE和部分纯化的胆道抗原Western blotting鉴定胆道上皮抗原，其中使用人胆道上皮细胞特异性的兔抗胆道抗原检测抗原。检测到≥200kd mw的B1、B2、B3抗原。它们分别从凝胶中电泳洗脱，用作刺激T细胞增殖的抗原。仅在PBC中检测到收缩1.0 ~ 0.1 mcg /ml的B1型细胞，在其他肝脏疾病中未检测到，而在PBC中未检测到B3型细胞。这些数据有力地指出了T细胞免疫对胆道上皮抗原的作用；B1抗原在PBC发病机制中的作用。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