Gruber R, Felber E, Pape G R, Höchtlen-Vollmar W, Riethmüller G
Institut für Immunologie, Universität München, Germany.
J Clin Lab Anal. 1994;8(5):284-92. doi: 10.1002/jcla.1860080507.
Antibodies directed against soluble liver antigen (SLA), liver kidney microsomal antigen (LKM-1-AG), and antimitochondrial antigen M2 (M2-AMA) are critical serological markers for the differential diagnosis of autoimmune chronic active hepatitis (AI-CAH) and primary biliary cirrhosis (PBC). The exact diagnosis of autoimmune hepatitis and PBC is of great clinical relevance, as it leads to different therapeutic strategies. In the present work, a simple and reliable ELISA test system is described, which applies the same test principle for the detection of three different species of autoantibodies important for the diagnosis of chronic liver disease. The ELISA assays are based on a competitive inhibition of binding of positive standard antibodies by patients sera containing antibodies of unknown specificity. The purified immunoglobulins of clinically and serologically clearly defined patients with SLA or LKM-1 positive AI-CAH and with M2 positive PBC were used as coating- and detection antibodies in the ELISAs. From homogenized rat liver the fractionated 100,000g supernatant was employed for the SLA ELISA, the microsomal preparation served as antigen for the LKM-1 ELISA and the mitochondrial preparation was used for the M2 ELISA. In 1,500 sera of patients with the differential diagnosis of a hepatobiliary disease, 17 gave a positive signal in the SLA-ELISA, 12 in the LKM-1-ELISA, and 72 in the M2-ELISA. The results of the ELISAs were compared with Western blotting and immunofluorescence staining pattern on cryostat sections and Hep2 cells. The antibody profiles of several patients are described in detail.
针对可溶性肝抗原（SLA）、肝肾微粒体抗原（LKM-1-AG）和抗线粒体抗原M2（M2-AMA）的抗体是自身免疫性慢性活动性肝炎（AI-CAH）和原发性胆汁性肝硬化（PBC）鉴别诊断的关键血清学标志物。自身免疫性肝炎和PBC的准确诊断具有重要的临床意义，因为这会导致不同的治疗策略。在本研究中，描述了一种简单可靠的ELISA检测系统，该系统采用相同的检测原理来检测对慢性肝病诊断重要的三种不同种类的自身抗体。ELISA检测基于含有未知特异性抗体的患者血清对阳性标准抗体结合的竞争性抑制。临床上和血清学上明确诊断为SLA或LKM-1阳性AI-CAH以及M2阳性PBC的患者的纯化免疫球蛋白用作ELISA中的包被抗体和检测抗体。从匀浆的大鼠肝脏中，100,000g分级上清液用于SLA ELISA，微粒体制备物用作LKM-1 ELISA的抗原，线粒体提取物用于M2 ELISA。在1500例患有肝胆疾病鉴别诊断的患者血清中，17例在SLA-ELISA中呈阳性信号，12例在LKM-1-ELISA中呈阳性，72例在M2-ELISA中呈阳性。将ELISA结果与冷冻切片和Hep2细胞上的蛋白质印迹和免疫荧光染色模式进行比较。详细描述了几位患者的抗体谱。
