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

抽象的。 这份申请是为了竞争性续期我们目前的R01资助，标题是 《改善免疫介导的牙周病》，共出版了23份相关出版物。 远远的。我们的研究主要集中在B10细胞在牙周病组织破坏中的抗炎作用 和骨吸收。具体地说，我们证明了B10细胞可以被激活和扩增 由特定的TLR信号和共刺激分子触发，从而抑制局部炎症和骨骼 实验性牙周炎在体内的丢失。观察到的抗炎作用与IL-10有关 B10细胞分泌。在证实这些细胞的抗炎作用的同时，B10的潜在作用 细胞在炎症消退中的作用在很大程度上是未知的。我们使用多光子活体显微镜的最新数据 单核/巨噬细胞与转移的B10细胞在牙周组织中的共存 诱发性牙周炎。在体外，活化的小鼠B10细胞中PD-L1的表达水平升高 脾细胞或人外周血单核细胞(PBMC)。与活化的B10细胞共培养的研究 单核细胞系(THP-1)来源的巨噬细胞上调巨噬细胞上PD-1的表达 巨噬细胞产生特殊的促分解介质(SPM)。这些影响被削弱了。 当B10与巨噬细胞的直接接触被阻断时。连同之前出版的 其他人的发现，我们假设i)激活的B10细胞诱导促分解巨噬细胞 通过PD-L1/PD-1结扎分化和产生专门的促分解介质(SPM)，以及 B10-巨噬细胞相互作用产生的SPM促进B10细胞分化的作用 发挥抗体分泌细胞和促巨噬细胞分解功能，促进炎症消退。 在这个方案中，我们将首先确定PD-L1/PD-1连接在B10介导的 B10-巨噬细胞与动物共培养时巨噬细胞表型转换及脂质介质的产生 过继B10细胞移植的实验性牙周炎模型(目标1)；然后SPM对B10细胞的作用 将确定B10-巨噬细胞相互作用过程中的分化以及它们对B细胞的影响。 介导病原体清除，并抑制牙周炎和牙周骨丢失(目标2)；第三， B10-巨噬细胞相互作用对巨噬细胞前分解功能、中性粒细胞活性的影响 将确定牙周炎症的消退(目标3)。这个项目的成功完成将 在理解免疫调节细胞-细胞相互作用及其相互作用方面产生概念上的进步 对炎症消退的影响。如果我们的假设是正确的，这些研究将拓宽我们对 免疫检查点分子PD-L1/PD-1在牙周炎症消退中的作用 疾病以及其他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”, which has produced a total of 23 related publications so far. Our studies have focused on the anti-inflammatory role of B10 cells in periodontal disease tissue destruction and 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 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. In vitro, the expression level of PD-L1 was elevated in activated B10 cells from mouse splenocytes or human peripheral blood mononuclear cells (PBMC). Co-culture of activated B10 cells with monocytic cell line (THP-1)-derived macrophages up-regulated PD-1 expression on macrophages and increased the production of specialized pro-resolving mediators (SPM) by macrophages. 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 that ii) 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. 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 animal model of experimental periodontitis with adoptive B10 cell transfer (Aim 1); Then the actions of 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); Thirdly, 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-cell interactions and their impact on the resolution of inflammation. If our hypothesis is correct, these studies will broaden our insights into the potential role of immune checkpoint molecules PD-L1/PD-1 in the resolution of inflammation in periodontal disease, as well as other immune-mediated osteolytic conditions such as osteoporosis or rheumatoid arthritis.