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采用免疫扩散法、对流免疫电泳法和免疫印迹法检测1型抗肝细胞溶质抗体(抗-LC1):不同技术的比较

Detection of anti-liver cytosol antibody type 1 (anti-LC1) by immunodiffusion, counterimmunoelectrophoresis and immunoblotting: comparison of different techniques.

作者信息

Muratori L, Cataleta M, Muratori P, Manotti P, Lenzi M, Cassani F, Bianchi F B

机构信息

Cattedra di Medicina Interna II, Università di Bologna, Policlinico S. Orsola, Italy.

出版信息

J Immunol Methods. 1995 Dec 1;187(2):259-64. doi: 10.1016/0022-1759(95)00192-x.

Abstract

Liver cytosol specific antibody type 1 (anti-LC1) was first described in a proportion of patients with liver/kidney microsomal antibody type 1 (anti-LKM1)-positive autoimmune hepatitis (AIH) and is routinely evaluated by immunodiffusion (ID). Using human liver cytosol as the source of antigen, we have used ID, counterimmunoelectrophoresis (CIE) and immunoblotting (IB), to test sera from 167 patients with documented chronic liver diseases of different etiology. 15 patients had antinuclear antibody (ANA) and/or smooth muscle antibody (SMA)-positive AIH, 13 had anti-LKM1-positive AIH, four had ANA/SMA/anti-LKM1-negative AIH, 76 had anti-LKM1-positive hepatitis C (recently renamed unclassified chronic hepatitis-UCH), 40 had chronic hepatitis C, 15 had chronic hepatitis B, and 4 had chronic hepatitis D. A precipitin line of identity with an anti-LC1 reference serum was detected both by ID and CIE in 16 patients: six with anti-LKM1-positive 'definite' AIH, four with ANA/SMA/anti-LKM1-negative 'definite' AIH, and six with anti-LKM1-positive UCH. By IB, 14 out of the 16 anti-LC1-positive sera (87.5%) reacted with a 58 kDa human liver cytosolic polypeptide, whereas three out of 16 (19%) recognised an additional 60 kDa band. Compared to ID, CIE is more economical in terms of both time and reagents and provides more clear-cut results. The 58 kDa reactivity by IB was detectable in nearly all CIE/ID anti-LC1-positive patients, was not found among CIE/ID anti-LC1-negative patients. In conclusion, CIE is the ideal screening test for the detection of anti-LC1, an autoantibody that can be regarded as an additional serological marker of AIH and is especially useful in ANA/SMA/anti-LKM1 negative cases.

摘要

肝细胞溶质特异性1型抗体(抗-LC1)最初在一部分1型肝肾微粒体抗体(抗-LKM1)阳性的自身免疫性肝炎(AIH)患者中被描述,并且通常通过免疫扩散(ID)进行评估。以人肝细胞溶质作为抗原来源,我们使用免疫扩散、对流免疫电泳(CIE)和免疫印迹(IB),检测了167例有不同病因慢性肝病记录患者的血清。15例患者抗核抗体(ANA)和/或平滑肌抗体(SMA)阳性的AIH,13例抗-LKM1阳性的AIH,4例ANA/SMA/抗-LKM1阴性的AIH,76例抗-LKM1阳性的丙型肝炎(最近重新命名为未分类慢性肝炎-UCH),40例慢性丙型肝炎,15例慢性乙型肝炎,4例慢性丁型肝炎。通过免疫扩散和对流免疫电泳在16例患者中检测到与抗-LC1参考血清具有同一性的沉淀线:6例抗-LKM1阳性的“明确”AIH,4例ANA/SMA/抗-LKM1阴性的“明确”AIH,以及6例抗-LKM1阳性的UCH。通过免疫印迹,16例抗-LC1阳性血清中的14例(87.5%)与一条58 kDa的人肝细胞溶质多肽发生反应,而16例中的3例(19%)识别出另一条60 kDa的条带。与免疫扩散相比,对流免疫电泳在时间和试剂方面更经济,并且提供更明确的结果。通过免疫印迹检测到的58 kDa反应性在几乎所有对流免疫电泳/免疫扩散抗-LC1阳性患者中均可检测到,在对流免疫电泳/免疫扩散抗-LC1阴性患者中未发现。总之,对流免疫电泳是检测抗-LC1的理想筛查试验,抗-LC1是一种自身抗体,可被视为AIH的另一种血清学标志物,在ANA/SMA/抗-LKM1阴性病例中尤其有用。

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