The Autoantibodies to α6β4 Integrin of Patients Affected by Ocular Cicatricial Pemphigoid Recognize Predominantly Epitopes Within the Large Cytoplasmic Domain of Human β41
The Autoantibodies to α6β4 Integrin of Patients Affected by Ocular Cicatricial Pemphigoid Recognize Predominantly Epitopes Within the Large Cytoplasmic Domain of Human β41
复制标题
眼部瘢痕性类天疱疮患者的 α6β4 整合素自身抗体主要识别人 β41 大细胞质域内的表位
作者：
K. Bhol;M. Dans;R. K. Simmons;C. Foster;F. Giancotti;A. Ahmed
本研究旨在探讨眼瘢痕性类天疱疮（OCP）患者自身抗体识别的抗原决定簇。OCP是一种主要影响结膜和其他粘膜的上皮下起泡性自身免疫性疾病。我们以前证明，一个cDNA克隆，分离的角质形成细胞表达文库，通过免疫亲和纯化的OCP自身抗体，编码β4整合素亚基的胞质结构域。我们随后的研究表明，所有OCP患者的血清都能识别人β4整联蛋白亚基。为了鉴定OCP的抗β4自身抗体的普遍表位，我们使用了用编码野生型β4亚基、无尾β4亚基或缺乏胞外结构域的β4亚基的载体转染的细胞系。未转染的细胞系用作对照。用OCP血清、来自OCP血清的IgG级分和免疫亲和纯化的OCP自身抗体分析来自这些细胞系的裂解物。将针对人β4整联蛋白的细胞外和细胞质结构域的Ab用作阳性对照，而将正常人血清和正常人IgG级分用作阴性对照。通过使用免疫印迹、免疫沉淀和FACS分析来确定OCP Ab的反应性。本研究的结果表明，OCP血清、OCP IgG组分和免疫亲和纯化的OCP自身抗体与人β4整联蛋白亚基的胞内而非胞外结构域反应。体外细胞培养实验证明OCP自身抗体与细胞质结合。这些研究结果的发病机制OCP的相关性进行了讨论。
This study was undertaken to characterize the antigenic determinants recognized by the autoantibodies of patients with ocular cicatricial pemphigoid (OCP). OCP is a subepithelial, blistering, autoimmune disease that mainly affects the conjunctiva and other mucous membranes. We previously demonstrated that a cDNA clone, isolated from a keratinocyte expression library by using immunoaffinity-purified OCP autoantibody, encoded the cytoplasmic domain of β4 integrin subunit. Our subsequent studies showed that sera from all the OCP patients that were tested recognize the human β4 integrin subunit. To identify the prevalent epitopes of the anti-β4 autoantibodies of OCP, we have used cell lines transfected with vectors encoding a wild-type β4 subunit, a tailless β4 subunit, or a β4 subunit lacking the extracellular domain. Nontransfected cell lines were used as controls. Lysates from these cell lines were analyzed with OCP sera, IgG fractions from OCP sera, and immunoaffinity-purified OCP autoantibodies. Abs to extracellular and cytoplasmic domains of human β4 integrin were used as positive controls, whereas normal human sera and normal human IgG fractions were used as negative controls. The reactivity of OCP Abs was determined by using immunoblotting, immunoprecipitation, and FACS analysis. The results of this study indicate that OCP sera, OCP IgG fractions, and immunoaffinity-purified OCP autoantibodies react with the intracellular and not the extracellular domain of human β4 integrin subunit. In vitro cell culture experiments demonstrated that OCP autoantibody binds to the cytoplasm of the cells. The relevance of these findings to the pathogenesis of OCP is discussed.
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作者：
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影响因子：
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DOI：
--
发表时间：
1994
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
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作者：
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Madaio,MP