Gene Expression Profiling of the Kveim Siltzbach Test

描述(由申请人提供)： 确定肉样肉芽肿形成早期基因表达的方法一直受到限制。目前尚不存在结节病的动物模型，已在诊断时或病程中获得的患者样本中进行了人体研究。之前的研究是在Mt.西奈半岛和其他地方的研究表明，对Kveim-Siltzbach(KS)混悬液的细胞和生化肉芽肿反应与疾病肉芽肿平行。对1、2、3和4周的KS注射组织的研究表明，CD4淋巴细胞和巨噬细胞的募集逐渐形成成熟的上皮样细胞、巨细胞和淋巴细胞边缘，形成结节样肉芽肿。在肉芽肿出现的这段时间内，没有关于基因表达的时间序列的研究。Kveim成熟过程中的基因表达微阵列分析为研究导致肉芽肿形成的一系列分子生物学事件提供了一个独特的机会。 我们建议招募5到8名最近KS试验呈阳性的患者来诊断他们的疾病。将进行四次皮下注射。在KS悬液孵育后1天、1周、2周和4周取材。作为对照，每个患者都将接受正常皮肤的活组织检查。基因表达谱将通过微阵列分析确定，并与组织学变化相关。基因芯片的基因表达结果将通过RT-PCR和免疫组织化学方法进行验证。识别KS悬浮液中表达的基因将提供有关肉芽肿形成的新信息，为未来的研究提供更大的方向，并可能有助于确定新的潜在干预靶点。

DESCRIPTION (provided by applicant): Approaches to define early gene expression in sarcoidal granuloma formation have been limited. No animal model for sarcoidosis exists and human studies have been carried out in patient samples obtained at diagnosis or during the disease course. Previous studies performed at Mt. Sinai and elsewhere have shown that the cellular and biochemical granulomatous response to Kveim-Siltzbach (KS) suspension parallels the disease granuloma. Studies of KS injected tissue harvested at weeks 1, 2, 3 and 4 have revealed recruitment of CD4 lymphocytes and macrophages that gradually form the mature epithelioid cells, giant cells, and rim of lymphocytes that form the sarcoidal granuloma. There are no studies of the temporal sequence of gene expression during this time period as granulomas emerge. Gene expression microarray analyses during Kveim maturation offer a unique opportunity to investigate the cascade of molecular biologic events leading to granuloma formation. We propose to recruit 5 to 8 patients who recently had positive KS tests for the diagnosis of their illness. Four subcutaneous injections will be performed. Sites will be biopsied a 1 day, 1 week, 2 weeks, and 4 weeks after incubation of KS suspension. For controls each patient will have a biopsy of normal skin. Gene expression profiles will be determined by microarray analysis and related to histological changes. Gene expression results by microarray will be verified by RT PCR and immunohistochemisty. Identifying genes expressed in response to the KS suspension will provide new information about the initiation of granuloma formation, give greater direction to future studies and could help identify new potential targets for intervention.