摘要
目的:观察扩张型心肌病(DCM)患者血清中是否独立存在抗心肌细胞L型钙离子通道(Human Cav1.2)自身抗体(简称:抗钙通道抗体)。方法:以合成多肽片段作为抗原,用ELISA法检测DCM患者(DCM组)及正常人(正常对照组)(各60例)血清中的抗钙通道抗体和抗心肌细胞线粒体内膜ADP/ATP载体蛋白(ANT)抗体。用亲合层析法提纯DCM患者(DCM组)血清中的抗钙通道抗体,以制备的大鼠抗Human Cav1.2α1亚单位多克隆抗体作为阳性对照,用免疫印记技术,免疫荧光组织化学技术检测DCM患者血清中抗钙通道抗体。结果:ELISA检测60例DCM患者中29例抗钙通道抗体阳性(29/60,阳性率48.33%),明显高于正常对照组(4/60,6.67%,P<0.01)。抗钙通道抗体和抗ANT抗体检出结果无一致性(P>0.05)。免疫印记中亲合层析法提纯的DCM患者抗钙通道抗体可特异地识别240000的多肽(Human Cav1.2α1c亚单位),结果与阳性对照相符。抗钙通道抗体经抗原充分吸附后特异性识别被阻断。免疫荧光实验显示,抗钙通道抗体能特异性地结合在大鼠心室肌细胞膜上。结论:DCM患者血清中存在新型的抗钙通道抗体。
Objective: In this study we sought to investigate whether there is an independent serum autoantibody against the cardiac L-type Ca^2+ channel (human Cav1. 2) in patients with dilated cardiomyopathy (DCM). Method:Peptide corresponding to the sequence (residues 2- 15) of the S1 segment, domain Ⅰ of 1 subunit, human Cavl. 2 was detected as an antigen to screen sera from patients with DCM (n= 60), and healthy blood donors (HBD, control group, n 60) in ELISA while the autoantibody against the adenine nucleotide translocator(ANT) was monitored. Affinity-purified autoantibodies from positive sera of patients with DCM and rat antipeptide polyclonal antibodies were deteceted by immunoblotting assay and immunofluorescence technique. Result:Sera from 29 patients with DCM (48. 33%) and 4 HBD (6.67%) recognized the peptide of human Cavl. 2 in ELISA and the difference was significant (P〈0.01). Autoantibody against the ANT and the autoantibody against the human Cavl. 2 were not in concordance. Affinity-purified autoantibody from positive sera of patients with DCM recognized a major band of about 240kD on the electrotransferred protein of rat ventricular cell membrane. This band was monospecific since its staining was blocked after preincubation with the human Cav1. 2 peptide and it coincided well with the band revealed by rat antipeptide polyclonal antibodies. In immunofluorescence, the stain was specially located with membranes of rat ventricular cardiocytes. Conclusion:These data confirme that there is a novel type of autoantibody against the human Cav1. 2 in the sera of patients with DCM.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2007年第8期601-605,共5页
Journal of Clinical Cardiology
关键词
心肌病
扩张型
钙通道
L型
自身抗体
Cardiomyopathy, dilated
Calcium Channels, L- Type
Autoantibodies