STEROID MECHANISMS IN CUTANEOUS ALLERGIC INFLAMMATION

本提案中的研究将利用皮肤过敏原激发， 确定观察到的糖皮质激素（GC）在 过敏性疾病的治疗。先前的研究表明， 全身和局部GC抑制细胞内流以及临床 过敏原激发后3 - 12小时观察到的反应参数， 这就是所谓的晚期反应（LPR）。细胞招募是一种高度 调节过程，需要协调内皮细胞粘附 分子（EAM）及其在循环白细胞上的反受体沿着 局部产生趋化性细胞因子。体外模型显示， GC能够抑制各种细胞系中细胞因子产生 即使在刺激之后。本提案中概述的研究将测试 GC抑制内皮细胞粘附分子表达假说 通过抑制已知的内皮激活细胞因子的产生 (IL-1、TNF、IL-4和干扰素-γ。本提案中的其他研究 将专注于一种新发现的趋化因子RANTES的调节， 已被证明是嗜酸性粒细胞的有效化学引诱剂， 记忆淋巴细胞，这两种细胞类型选择性地招募， LPR。为了验证这些假设，我们将采用常规组织学， 免疫组织化学和原位杂交方法， 过敏性受试者在实验性过敏原攻击后的活检。在 此外，将通过以下方法处理活检样本以分析细胞因子mRNA： RNA酶保护试验和ELISA用于检测RANTES和可溶性 黏附分子 这项工作提供的信息可能有助于我们更好地了解 GC缓解过敏性疾病的机制以及其他 炎性病症如结缔组织疾病，血管炎， 炎症性皮肤病等。更好地了解GC如何提供 治疗这些疾病的益处可能最终导致 具有GC功效的抗炎药物的工程化 减少副作用。

Studies in this proposal will utilize cutaneous allergen challenge to determine the basis for the observed efficacy of glucocorticoids (GC) in the treatment of allergic diseases. Previous studies have shown that both systemic and topical GC inhibit the cellular influx as well as the clinical parameters of the reaction seen 3 - 12 hours after allergen challenge, the so-called late phase reaction (LPR). Cellular recruitment is a highly regulated process that requires the coordination of endothelial adhesion molecules (EAM) and their counterreceptors on circulating leukocytes along with local production of chemotactic cytokines. In vitro models have shown that GC are capable of inhibiting cytokine production in various cell lines even after stimulation. Studies outlined in this proposal will test the hypothesis that GC inhibit the expression of endothelial adhesion molecules by inhibiting the production of the known endothelial-activating cytokines (IL-1, TNF, IL-4, and interferon-gamma. Additional studies in this proposal will focus on the regulation of a newly-identified chemokine called RANTES, which has been shown to be a potent chemoattractant for eosinophils and memory lymphocytes, two cell types that are selectively recruited in the LPR. To test these hypotheses we will employ routine histology, immunohistochemistry, and in situ hybridization methodologies using biopsies of allergic subjects following experimental allergen challenge. In addition, biopsy samples will be processed for analysis of cytokine mRNA by RNase protection assays and ELISAs for detection of RANTES and soluble adhesion molecule. Information provided by this work may help us to better understand the mechanisms by which GC alleviate allergic diseases as well as other inflammatory conditions such as connective tissue diseases, vasculitides, inflammatory dermatoses, etc. A better understanding of how GC provides benefit in the treatment of such disorders may eventually lead to the engineering of an anti-inflammatory medication which has the efficacy of GC with reduced side effects.