ISOLATION OF A HUMAN EPIDERMAL CDNA CORRESPONDING TO THE 180-KD AUTOANTIGEN RECOGNIZED BY BULLOUS PEMPHIGOID AND HERPES-GESTATIONIS SERA - IMMUNOLOCALIZATION OF THIS PROTEIN TO THE HEMIDESMOSOME
ISOLATION OF A HUMAN EPIDERMAL CDNA CORRESPONDING TO THE 180-KD AUTOANTIGEN RECOGNIZED BY BULLOUS PEMPHIGOID AND HERPES-GESTATIONIS SERA - IMMUNOLOCALIZATION OF THIS PROTEIN TO THE HEMIDESMOSOME
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DOI：
10.1172/jci114812
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发表时间：
1990-10-01
影响因子：
15.9
通讯作者：
GIUDICE, GJ
中科院分区：
文献类型：
--
作者：
DIAZ, LA;RATRIE, H;GIUDICE, GJ
大疱性类天疱疮（BP）患者血清中存在的自身抗体与正常人皮肤的基底膜区结合，通常识别两种表皮蛋白，即BP 240和BP 180抗原。基于与BP血清的反应性，已经鉴定了来自λ gt 11人角质形成细胞文库的两个BP抗原cDNA克隆。单克隆抗体产生的融合蛋白（FP）可特异性地识别BP 180抗原，免疫印迹分析显示与BP 240无交叉反应。第二克隆产生的FP免疫吸附与BP 240表皮抗原特异性反应的自身抗体。北方印迹分析表明，BP 180和BP 240抗原分别由长度为6.0和8.5 kb的不同RNA转录本编码。BP 180溶原提取物的免疫印迹分析鉴定了135-kD FP，其被16份BP血清中的7份和8份妊娠疱疹血清中的7份识别。兔抗溶原性BP 180 FP的抗血清通过免疫印迹特异性识别人表皮提取物中的BP 180抗原，通过间接免疫荧光标记BMZ，并通过免疫电镜结合人表皮半桥粒。这些结果表明，BP和妊娠疱疹自身抗体识别的BP 180抗原是一种独特的半桥粒多肽，与BP 240抗原不同。
Autoantibodies present in the sera of patients with bullons pemphigoid (BP) bind to the basement membrane zone of normal human skin and commonly recognize two epidermal proteins, the BP240 and BP180 antigens. Two BP antigen cDNA clones from a lambda gt11 human keratinocyte library have been identified on the basis of reactivity with a BP serum. The fusion protein (FP) produced by one clone immunoadsorbed autoantibodies, which specifically recognized the BP180 antigen, showing no cross-reactivity with BP240 by immunoblot analysis. The FP produced by the second clone immunoadsorbed autoantibodies which specifically reacted with the BP240 epidermal antigen. Northern blot analysis demonstrated that the BP180 and BP240 antigens are encoded by distinct RNA transcripts with lengths of 6.0 and 8.5 kb, respectively. Immunoblot analysis of the BP180 lysogen extract identified a 135-kD FP which was recognized by 7 of 16 BP sera and 7 of 8 herpes gestationis sera. A rabbit antiserum prepared against the lysogenic BP180 FP specifically recognized the BP180 antigen from human epidermal extracts by immunoblotting, labeled the BMZ by indirect immunoflorecence, and bound to human epidermal hemidesmosomes by immuno-electron microscopy. These results indicate that the BP180 antigen recognized by BP and herpes gestationis autoantibodies is a unique hemidesmosomal polypeptide, distinguishable from the BP240 antigen.