Gene Expression Profiling of the Kveim Siltzbach Test

描述（由申请人提供）： 确定结节性肉芽肿形成早期基因表达的方法有限。目前还没有结节病的动物模型，在诊断时或病程中获得的患者样本中进行了人体研究。以前在Mt.西奈半岛和其他地方已经表明，对Kveim-Siltzbach（KS）悬浮液的细胞和生化肉芽肿反应与疾病肉芽肿相似。在第1、2、3和4周收获的KS注射组织的研究已经揭示了CD 4淋巴细胞和巨噬细胞的募集，其逐渐形成成熟的上皮样细胞、巨细胞和淋巴细胞的边缘，形成结节样肉芽肿。在肉芽肿出现的这段时间内，没有研究基因表达的时间顺序。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分子生物学事件级联提供了独特的机会。 我们建议招募5至8名最近KS试验阳性的患者，以诊断他们的疾病。将进行4次皮下注射。将在KS混悬液孵育后1天、1周、2周和4周对部位进行活检。对于对照组，每例患者将进行正常皮肤活检。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和免疫组化进行验证。鉴定对KS悬浮液反应的基因将提供有关肉芽肿形成起始的新信息，为未来的研究提供更大的方向，并有助于鉴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.