CHEMOKINE RANTES AND ALLERGIC INFLAMMATION

几个实验室在过去十年里的研究强烈表明， 嗜酸性粒细胞在哮喘的病理生理学中起着重要作用， 其他过敏性疾病。 这些细胞似乎是有选择性地招募 过敏反应的部位，并产生介质， 解释了这些反应的许多组织病理学特征。 的 本提案的总体目标是促进我们对 嗜酸性粒细胞在变应性炎症中的募集机制 研究嗜酸性粒细胞募集趋化因子RANTES在 过敏原诱导的炎症的不同体内模型和研究 糖皮质激素对体内外RANTES产生的影响。 具体目标：1. 为了评估RANTES是否在体内产生， 在皮肤中实验性抗原激发后的过敏受试者 或者肺 a）从收集的皮肤组织中提取总RNA 抗原激发后长达24小时，或来自活检样本， 抗原处理后19小时，来自肺的支气管肺泡灌洗液细胞 激发过敏性受试者以确定 RANTES。 将使用RNA酶保护试验检测RANTES mRNA，或 如果需要，通过半定量聚合酶链反应（PCR）。 RANTES 将通过进行特异性ELISA测定来确认体内生成 BAL液和皮肤或肺组织提取物。 的比较 非过敏或对照抗原激发，生理盐水激发过敏 个人将被执行。 2. 为了验证这个假设， 糖皮质激素在体内和体外抑制RANTES的产生。 因为在 体内研究中，糖皮质激素预处理将测试其 在先前描述的体内模型中对RANTES产生的影响。 在体外实验中，已知产生RANTES的细胞类型 组成型和活化后（包括THP-1， 横纹肌肉瘤细胞、人滑膜成纤维细胞和凝集素刺激的 人淋巴细胞）将暴露于糖皮质激素长达24小时 在刺激之前，RANTES mRNA以及成熟蛋白将 进行分析。 本提案中所述的实验结果 希望能帮助我们进一步了解 过敏性炎症和抗炎作用的机制 糖皮质激素

Studies by several laboratories over the last ten years strongly indicate that eosinophils play a central role in the pathophysiology of asthma and other allergic diseases. These cells appear to be selectively recruited to the sites of allergic reactions and them produce mediators which account for many of the histopathologic features of these reactions. The overall goal of this proposal is to advance our understanding of the mechanism of eosinophil recruitment during allergic inflammation by investigating the role of the eosinophil-recruiting chemokine RANTES in different in vivo models of allergen-induced inflammation and the study the effects of glucocorticoids on RANTES production in vitro and in vivo. Specific aims: 1. To assess whether RANTES is produced in vivo in allergic subjects following experimental antigen challenge in the skin or the lungs. a) Total RNA will be extracted from skin tissue collected up to 24hr after antigen challenge, or from biopsy samples and bronchoalveolar lavage cells derived from the lungs 19 hr after antigen challenge of allergic subjects to determine the presence of mRNA for RANTES. RANTES mRNA will be detected using an RNase protection assay or by semiquantitative polymerase chain reaction (PCR) if required. RANTES generation in vivo will be confirmed by a specific ELISA assay performed on BAL fluids and extracts of skin or lung tissue. A comparison of antigen challenge of non-allergic or control, saline-challenged allergic individuals will be performed. 2. To test the hypothesis that glucocorticoids inhibit RANTES production in vivo and in vitro. For in vivo studies, pretreatment with glucocorticoids will be tested for its effect on RANTES generation in the in vivo models previously described. In the in vitro experiments, cell types known to generate RANTES both constitutively and following activation (including THP-1, rhabdomyosarcoma cells, human synovial fibroblasts and lectin-stimulated human lymphocytes) will be exposed to a glucocorticoid for up to 24 hr prior to stimulation and RANTES mRNA, as well as the mature protein, will be assayed. The results of the experiments described in this proposal will hopefully help advance our knowledge of the pathophysiologic mechanisms of allergic inflammation and of the anti-inflammatory actions of glucocorticoids.