Calprotectin-mediated CD69 signaling in periodontitis

7.项目概要/摘要 牙周炎是一种慢性炎症性疾病，其特征在于牙周组织的破坏， 成年人牙齿脱落的主要原因。它是由一个生态失调的微生物生物膜，殖民牙龈沟。 我们试图确定和表征局部免疫反应的微生物生物膜，导致牙周炎。 最近，据报道，CD69在T调节细胞上的接合诱导免疫抑制活性。一 用于CD69介导的T细胞活化的天然配体是钙卫蛋白（CLP;与S100A9复合的S100A8; S100A8/A9; MRP8/14）。当在复层鳞状上皮中表达时，这种二价阳离子结合复合物 似乎有助于上皮内抗菌防御。当从感染或脱皮中释放出来时 然而，CLP可能与CD69 + T调节细胞或T辅助细胞17相互作用，最终 抑制免疫反应如果是这样的话，CLP可能会在启动期间发挥作用， 这与其作为促炎性"警报素"的假定功能相反。使用全局CLP空鼠标， 我们的初步数据表明，CLP的净效应抑制了急性炎症浸润的募集 并限制结扎诱导的实验性牙周炎模型中的牙周骨破坏。我们现在将 探索牙龈卟啉单胞菌（Porphyromonas gingivalis，Pg）致敏的小鼠结扎性牙周炎模型。 在解决问题的阶段 我们假设CLP信号通过CD69在启动过程中， 和解决阶段 实验 牙周炎，以抑制牙龈中的破坏性细胞浸润。为了验证我们的假设，我们 将：1：表征初始治疗期间可归因于CLP的炎性细胞浸润的差异 和解决阶段 实验性牙周炎2：确定Pg致敏的Treg细胞如何调节 通过CD69信号传导募集初始炎性细胞浸润 还有CLP。3：确定Pg致敏的Thl7细胞对调节初始Thl7细胞的募集的贡献。 和解决阶段 和解决阶段 炎症细胞浸润可归因于CD69信号传导和CLP。对我们 知识，我们是第一个有数据表明CLP抑制先天免疫反应的小组。 CD69依赖性。我们有工具来解释CLP如何有助于先天免疫的招募， 通过使用牙周炎的鼠模型在体内影响Treg和Th17细胞来治疗牙周炎。最终，我们将 表征Treg和Th17细胞中的CLP和CD69信号传导如何塑造免疫细胞环境， 牙龈驱动器保护牙周组织或破坏牙槽骨。获得的结果 本文将用于设计针对增强或抑制CLP活性的治疗干预。关键 将确定可能适合于人类靶向治疗干预的步骤。最终我们 旨在减轻牙周病的经济及个人负担。

7. PROJECT SUMMARY / ABSTRACT Periodontitis is a chronic inflammatory condition characterized by the destruction of the periodontium and is the leading cause of tooth loss in adults. It is driven by a dysbiotic microbial biofilm that colonizes the gingival sulcus. We seek to identify and characterize the local immune response to the microbial biofilm that leads to periodontitis. Recently, CD69 engagement on T regulatory cells was reported to induce immunosuppressive activities. A natural ligand for CD69-mediated activation of Tregs is calprotectin (CLP; S100A8 complexed to S100A9; S100A8/A9; MRP8/14). When expressed in stratified squamous epithelia, this divalent cation-binding complex appears to contribute to intraepithelial antimicrobial defense. When released from infected or desquamating keratinocytes or neutrophils, however, CLP may interact with CD69+ T regulatory or T helper 17 cells, ultimately suppressing the immune response. If so, CLP may function during the initiation of periodontitis contrary to its postulated function as a proinflammatory “alarmin”. Using a global CLP null mouse, our preliminary data suggest that the net effect of CLP dampens the recruitment of an acute inflammatory infiltrate and limits periodontal bone destruction in a ligature-induced experimental periodontitis model. We will now explore a modified mouse model of ligature-induced periodontitis primed with Porphyromonas gingivalis (Pg). and in the resolution phase We hypothesize that CLP signals through CD69 during the initiation and resolution phases of experimental periodontal inflammation to dampen the destructive cellular infiltrate in the gingiva. To test our hypothesis, we will: 1: Characterize the differences in the inflammatory cell infiltrate attributable to CLP during the initial and resolution phases of experimental periodontitis. 2: Determine how Pg-primed Treg cells modulate the recruitment of the initial inflammatory cell infiltrates through CD69 signaling and CLP. 3: Determine the contribution of Pg-primed Th17 cells to modulating recruitment of the initial and resolution phase and resolution phase inflammatory cell infiltrates attributable to CD69 signaling and CLP. To our knowledge, we are the first group with data suggesting that CLP dampens the innate immune response in a CD69-dependent manner. We have the tools to explain how CLP contributes to recruitment of innate immune cells by affecting Treg and Th17 cells in vivo using a murine model of periodontitis. Ultimately, we will characterize how CLP and CD69 signaling in Treg and Th17 cells shapes the immune cell environment in the gingiva to drives either protection of periodontal tissues or destruction of alveolar bone. The results obtained here will be used to design therapeutic interventions directed at boosting or inhibiting the activity of CLP. Critical steps will be identified that might be amenable to targeted therapeutic intervention in humans. Ultimately we aim to reduce the economic and personal burden of periodontal diseases.