Identification and Regulation of Defensins in Human Mucosal Tissue

正如我们在最初的申请中所述，鉴于新的证据表明人类β防御素（hBD）是重要的， 作为先天免疫和适应性免疫之间的免疫调节桥梁，我们有必要了解 这些肽在广泛的人类粘膜组织中的调节。它们是否受到同样的监管， 不同的粘膜？它们在上皮组织的各种疾病表现中是否受到相同的调节？到 解决这些问题，我们将完善我们的知识，防御素方面，他们的表达在各种 并更仔细地检查它们在各种粘膜部位内表达的调节。我们 因此，建议本提案的目标如下： 具体目标一。在健康和患病口腔中进行hBD-2和hBD-3的免疫定位研究， 皮肤和肠粘膜。 比较hBD-2和hBD-3在正常口腔、肠道和皮肤上皮中的免疫定位。 确定是否防御素表达或定位的变化在上皮疾病的各种 粘膜组织 具体目标二。研究hBD-2和hBD-3在口腔、皮肤和口腔粘膜中表达的调节机制。 肠上皮和细胞。 确定原代和永生化的口腔、肠道和皮肤上皮细胞是否上调hBD-2和hBD-3 响应于已知的防御素表达诱导剂，包括IL-22和IL-17的组合。 使用从以下获得的原代细胞确定防御素上调的反应是否被修饰： 相应粘膜的组织疾病（例如天疱疮、IBD、角化过度）。 检查天然防御素诱导剂（如EGF，TGF α，IGF-1）的表达和共定位 免疫组织化学分析。 确定促炎性TLR激动剂和免疫调节hBD-2和-3表达 肠上皮细胞分泌的细胞因子。

As stated in our original application, given the new evidence that human (3 defensins (hBDs) are important immunomodulatory bridging agents between innate and adaptive immunity, it behooves us to understand the regulation of these peptides in a broad range of human mucosal tissue. Are they regulated the same in different mucosae? Are they regulated the same in various disease manifestations of epithelial tissue? To address these questions we will refine our knowledge of defensins with regard to their expression in various mucosae and to more closely examine the regulation of their expression within various mucosal sites. We therefore propose the following aims for this proposal: Specific Aim I. Conduct immunolocalization studies of hBD-2 and hBD-3 in healthy and diseased oral, skin and intestinal mucosae. ¿ Compare the immunolocalization of hBD-2 and hBD-3 in normal oral, gut and skin epithelium. ¿ Determine if defensin expression or localization changes within epithelial disease in the various mucosal tissue. Specific Aim II. Examine the regulatory mechanism for expression of hBD-2 and hBD-3 in oral, skin and intestinal epithelium and cells. ¿ Determine if primary and immortalized oral, gut and skin epithelial cells upregulate hBD-2 and hBD-3 in response to known inducers of defensin expression, including the combination of IL-22 and IL-17. Determine if the response of defensin upregulation is modified using primary cells obtained from tissue disorders for the respective mucosae (eg, Pemphigus, IBD, hyperkeratosis). ¿ Examine expression and co-localization of natural defensin-inducing agents (eg, EGF, TGFa, IGF-1) using immunohistochemical analysis. ¿ Determine the regulation of hBD-2 and -3 expression by proinflammatory TLR-agonists and immune cytokines by intestinal epithelial cells.