Yamamoto A M, Johanet C, Duclos-Vallee J C, Bustarret F A, Alvarez F, Homberg J C, Bach J F
Service d'Immunologie, Hôpital Necker, Paris, France.
Clin Exp Immunol. 1997 Jun;108(3):396-400. doi: 10.1046/j.1365-2249.1997.4071302.x.
Antibodies specific for cytochrome CYP2D6, formally known as liver-kidney-microsome type-1 antibodies (LKM-1), are characteristically found in a subgroup of patients presenting autoimmune hepatitis. They are also found in some patients with chronic HCV infection. These autoantibodies are usually detected by indirect immunofluorescence, immunoblotting and ELISA tests. In an attempt to set up a more sensitive detection assay we developed a quantitative immunoprecipitation radioligand assay using a 35S-methionine-labelled CYP2D6 antigen obtained by in vitro transcription and translation synthesis. All 16 sera from AIH-2 patients strongly bound to this CYP2D6 antigen. Two of the nine sera (22%) from AIH-2 patients that presented only liver cytosol-1 antibodies also bound to CYP2D6. All 24 sera from HCV patients that were positive for LKM-1 antibodies by indirect immunofluorescence were also positive using this CYP2D6 radioligand assay. Lastly, all 15 sera from HCV patients negative for LKM-1 antibodies were negative by this test. The present results support the view that this quantitative radioligand assay is more sensitive than immunoblotting and ELISA CYP2D6 assays, and that it could be used in combination with indirect immunofluorescence assay.
细胞色素CYP2D6特异性抗体,以前称为肝肾微粒体1型抗体(LKM-1),在自身免疫性肝炎患者的一个亚组中具有特征性发现。它们也在一些慢性丙型肝炎病毒(HCV)感染患者中被发现。这些自身抗体通常通过间接免疫荧光、免疫印迹和酶联免疫吸附测定(ELISA)检测。为了建立一种更灵敏的检测方法,我们开发了一种定量免疫沉淀放射性配体测定法,使用通过体外转录和翻译合成获得的35S-甲硫氨酸标记的CYP2D6抗原。所有16份自身免疫性肝炎2型(AIH-2)患者的血清都与这种CYP2D6抗原强烈结合。9份仅呈现肝细胞溶质1抗体的AIH-2患者血清中有2份(22%)也与CYP2D6结合。通过间接免疫荧光检测LKM-1抗体呈阳性的所有24份HCV患者血清,使用这种CYP2D6放射性配体测定法也呈阳性。最后,15份LKM-1抗体呈阴性的HCV患者血清通过该检测均为阴性。目前的结果支持这样的观点,即这种定量放射性配体测定法比免疫印迹和ELISA CYP2D6测定法更灵敏,并且它可以与间接免疫荧光测定法联合使用。