PATHOGENIC ROLE OF RO/SS-A AUTOANTIGENS IN SKIN DISEASE

RO/SS-A 自身抗原在皮肤病中的致病作用

基本信息

批准号：
2006253
负责人：
DANIEL P MCCAULIFFE
金额：
$ 10.64万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-12-01 至 1998-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2006253
关键词：
autoantigens cellular pathology confocal scanning microscopy discoid lupus erythematosus fluorescence microscopy human subject intracellular transport keratinocyte laboratory mouse laboratory rabbit monoclonal antibody recombinant proteins skin disorder tissue /cell culture ultraviolet radiation

项目摘要

Several lines of evidence indicate that the Ro/SS-A (Ro) autoantibodies and their target autoantigens play major roles in the pathogenesis of subacute cutaneous lupus erythematosus (SCLE) and neonatal LE (NLE). Recently cDNA clones have been characterized that encode immunologically distinctive 52 kD and 60 kD Ro autoantigens which share no significant amino acid sequence homology. These cDNA clones will be utilized in the proposed studies aimed at further elucidating the role that these autoantigens and autoantibodies play in the pathogenesis of SCLE and NLE skin disease. The specific short term goals of this project are to: 1. Develop a panel of antibodies that are specific for the 52 and 60 kD Ro molecules. To accomplish this, Ro antibodies will be affinity purified from anti-Ro rabbit antisera and Ro autoimmune sera by using recombinant human 52 or 60 kD Ro protein. Murine monoclonal antibodies will also be raised against each recombinant Ro protein. The development of 52 and 60 kD Ro specific antibodies is of paramount importance as these antibodies will allow investigators to more precisely study each Ro autoantigen. These antibodies will serve as valuable tools not only for the proposed studies but for future studies as well. 2. Determine the effects of clinically-relevant perturbations on human keratinocyte expression and intracellular distribution of the Ro autoantigens. These studies will be done on cultured human keratinocytes and in both normal human skin and SCLE patient skin. Ro specific antibodies will detect their respective antigens by microscopy (conventional immunofluorescence, scanning confocal, and immunoelectron). Ro protein expression will be quantified by ELISA inhibition and whole cell ELISA methods. Knowledge gained from these studies should further elucidate the roles that the Ro autoantigens and autoantibodies play in the pathogenesis of SCLE and NLE, and provide a new foundation on which future investigative studies can be devised. It is hoped that these efforts will provide information that serves to expedite the development of more effective therapies for these LE-related skin diseases.

几条证据表明RO/SS-A（RO）自身抗体他们的目标自动抗原在亚急性皮肤红斑狼疮（SCLE）和新生儿LE（NLE）。最近已经表征了cDNA克隆，该克隆在免疫学上编码独特的52 kd和60 kd RO自动抗原，没有显着的氨基酸序列同源性。这些cDNA克隆将在拟议的研究旨在进一步阐明这些作用自动抗原和自身抗体在SCLE和NLE的发病机理中发挥作用皮肤病。该项目的具体短期目标是： 1。开发一个针对52和60 kD RO的抗体面板分子。为此，RO抗体将是亲和力纯化的通过使用重组抗Ant-Ro Rabbit Antisera和RO自身免疫性血清人52或60 kD RO蛋白。鼠单克隆抗体也将是针对每个重组RO蛋白提出。 52和60的发展 KD RO特异性抗体至关重要，因为这些抗体将允许研究人员更精确地研究每个RO自动抗原。这些抗体不仅可以作为提议的有价值的工具研究，但也用于未来的研究。 2。确定与临床相关的扰动对人的影响 RO的角质形成细胞表达和细胞内分布自动抗原。这些研究将对培养的人角质形成细胞进行在正常的人皮肤和SCLE患者皮肤中。特定于ro 抗体将通过显微镜检测其各自的抗原（常规的免疫荧光，扫描共聚焦和免疫电子）。 RO蛋白表达将通过ELISA抑制和整体量化细胞ELISA方法。从这些研究中获得的知识应进一步阐明角色 RO自动抗原和自身抗体在 Scle和NLE，并为未来的调查提供了一个新的基础可以设计研究。希望这些努力能够提供有助于加快开发更有效的信息这些与LE相关的皮肤病的疗法。