Regulatory B Cell in the Amelioration of Immune-Mediated Periodontal Disease

抽象的。 本申请是为了我们目前的R 01资助的竞争性更新，标题为“调节B细胞在 免疫介导的牙周病的改善”。我们目前的研究集中在抗- B10细胞在牙周病组织破坏和炎性骨吸收中炎症作用 具体地说，我们证明了B10细胞可以在特定TLR触发后活化和扩增， 信号传导和共刺激分子，从而抑制实验性骨丢失中的局部炎症和骨丢失。 体内牙周炎。观察到的抗炎作用与B10细胞分泌IL-10有关。 虽然证实了这些细胞的抗炎作用，但B10细胞在解决炎症反应中的潜在作用仍然存在。 炎症的阶段基本上是未知的。我们最近使用多光子活体显微镜的数据表明， LPS诱导的牙龈炎的牙龈组织中单核细胞/巨噬细胞与转移的B10细胞的共定位。 来自人外周血单核细胞（PBMC）的活化的B10细胞与人单核细胞的共培养 细胞系（THP-1）分化的巨噬细胞上调巨噬细胞的PD-1表达， 专业的亲解决调解员（SPM）。当B10和B20之间直接接触时， 巨噬细胞被阻断。结合其他人先前发表的研究结果，我们假设， i）活化的B10细胞诱导促消退巨噬细胞分化和特化促消退巨噬细胞分化， ii）通过PD-L1/PD-1连接介导物（SPM）产生，和ii）来源于B10- 巨噬细胞相互作用增强B10细胞向抗体分泌细胞的分化， 分解巨噬细胞功能，促进炎症消退，减轻牙周骨丢失。 在这个建议中，我们将首先确定PD-L1/PD-1连接对B10介导的调节的作用。 巨噬细胞表型转换和使用B10-巨噬细胞共培养物产生脂质介质， 过继性B10细胞转移（目的1）;特异性促消退介质（SPM）对B10的作用 将确定B10-巨噬细胞相互作用期间的分化，以及它们对B细胞的作用。 介导的病原体清除，以及抑制牙龈炎症和牙周骨丢失（目的2）;最后， B10-巨噬细胞相互作用对促消退巨噬细胞功能、中性粒细胞活性和随后的 将确定牙周炎症的消退（目标3）。该项目的成功完成将 在我们对免疫调节细胞相互作用及其影响的理解中产生概念上的进步 炎症的消退。如果我们的假设是正确的，这些研究将拓宽我们的视野， 免疫检查点分子PD-L1/PD-1在牙周病炎症消退中的作用，以及 其它免疫介导的溶骨性疾病，如骨质疏松症或类风湿性关节炎。

Abstract. This application is for the competitive renewal of our current R01 grant titled “Regulatory B cells in the amelioration of immune-mediated periodontal disease”. Our current studies have focused on the anti- inflammatory role of B10 cells in periodontal disease tissue destruction and inflammatory bone resorption. Specifically, we demonstrated that B10 cells can be activated and expanded following triggering by specific TLR signaling and co-stimulatory molecules and thereby inhibit local inflammation and bone loss in experimental periodontitis in vivo. The observed anti-inflammatory effects are associated with IL-10 secretion by B10 cells. While substantiating the anti-inflammatory actions of these cells, the potential role of B10 cells in the resolution phase of inflammation is largely unknown. Our recent data using multiphoton intravital microscopy demonstrated co-localization of monocytes/ macrophages with transferred B10 cells in gingival tissue of LPS-induced gingivitis. Co-culture of activated B10 cells from human peripheral blood mononuclear cells (PBMC) with human monocytic cell line (THP-1)-differentiated macrophages up-regulated PD-1 expression by macrophages and production of specialized pro-resolving mediators (SPM). These effects were diminished when direct contact between B10 and macrophages was blocked. Together with the previously published findings by others, we hypothesize that i) activated B10 cells induce pro-resolving macrophage differentiation and specialized pro-resolving mediators (SPM) production via PD-L1/PD-1 ligation, and ii) that actions of SPM derived from B10- macrophage interaction enhance both B10 cell differentiation to antibody-secreting cells and pro- resolving macrophage function to promote inflammation resolution and alleviate periodontal bone loss. In this proposal, we will first determine the role of PD-L1/PD-1 ligation on B10-mediated regulation of macrophage phenotype switch and production of lipid mediators using B10-macrophage co-cultures and adoptive B10 cell transfer (Aim 1); The actions of specialized pro-resolving mediators (SPM) on B10 differentiation during B10-macrophage interaction will be determined, together with their effects on B cell- mediated pathogen clearance, and inhibition of gingival inflammation and periodontal bone loss (Aim 2); Lastly, B10-macrophage interaction on pro-resolving macrophages function, neutrophil activity, and the subsequent resolution of periodontal inflammation will be determined (Aim 3). Successful completion of this project will generate conceptual advances in our understanding of the immune regulatory cell interactions and their impact on the resolution of inflammation. If our hypothesis is correct, the studies will broaden our insights into possible role of immune checkpoint molecules PD-L1/PD-1 in inflammation resolution in periodontal disease, as well as other immune-mediated osteolytic conditions such as osteoporosis or rheumatoid arthritis.