Dendritic Cells and Periodontal Disease

树突状细胞和牙周病

• 批准号: 8664246
• 负责人: DANA T GRAVES
• 金额: $ 40万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8664246
• 关键词: Address; Alveolar Bone Loss; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Bacteria; Binding; Biological Assay; Bone Resorption; Breeding; Calvaria; Cells; DNA; DNA Binding; Data; Dendritic Cells; Disease Progression; Down-Regulation; Event; Exhibits; Flow Cytometry; Gene Expression; Gene Targeting; Generations; Genetic Transcription; Goals; Histologic; ITGAX gene; Immune response; Immunization; In Vitro; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Interferons; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-6; Link; Measurement; Measures; Mediating; Modeling; Monitor; Mus; Nuclear; Oral; Oral cavity; Osteoclasts; Pathogenesis; Periodontal Diseases; Periodontium; Play; Porphyromonas gingivalis; Process; Production; Promoter Regions; Regulatory Pathway; Reporting; Role; Scalp structure; Signal Transduction; Site; Spleen; TNF gene; Testing; Up-Regulation; base; bone; bone loss; chromatin immunoprecipitation; cytokine; in vivo; in vivo Model; interleukin-12 subunit p40; mutant; oral infection; osteoclastogenesis; pathogen; prevent; promoter; recombinase; research study; response; soft tissue; transcription factor

DESCRIPTION (provided by applicant): Dendritic cells (DC) may play a central role in the pathogenesis of periodontal disease by participating in events that link bacterial stimulation to periodontal bone loss. They may accomplish this by the production of factors that regulate the adaptive and innate immune responses. Furthermore, immature DCs have been reported to function as osteoclasts precursors. Preliminary Data examining DCs in vitro establish that the transcription factor FOXO1 is essential for LPS up-regulation of several cytokines, IL-1, TNF, IL-6, and IL-12 and down-regulation of the anti-inflammatory cytokine IL-10. Moreover, Akt plays an important role in this process by reducing FOXO1 nuclear localization and limiting inflammatory cytokine production in DC. Thus, we propose that FOXO1 is critical in upregulating an inflammatory response in DCs and that a hyper- inflammatory response is prevented by Akt. These findings serve as the basis for the current proposal in which we will test in vivo the hypothesis that the FOXO1-Akt axis regulates DC cytokine expression. Moreover, we will determine whether this regulatory pathway is essential for stimulating the adaptive immune response to the periodontal pathogen, P. gingivalis, and whether it plays a significant role in bacteria induced bone loss. To delete FOXO1 in DCs we will use the Cre-lox approach. We have already bred floxed FOXO1 mice with mice that express Cre recombinase under control of the CD11c promoter. DC isolated from the resulting experimental (CD11cCre+/FOXO1L/L) mice exhibit reduced cytokine expression stimulated by LPS compared to littermate control mice (CD11cCre-/FOXO1L/L). We will examine host-bacteria interactions in vivo by injecting P. gingivalis in the calvarial model. Because mice are naive to P. gingivalis we can examine the response when the adaptive immune response is not present and compare it to mice in which the adaptive immune response is activated by pre-immunization with P. gingivalis. By flow cytometry we will have a detailed analysis on the impact of FOXO1 deletion in generating a systemic and local adaptive immune response. The underlying calvarial bone will also be examined histologically to investigate the subsequent effect on osteoclastogenesis and bone resorption. The oral gavage model of applying P. gingivalis to the oral cavity will be studied in experimental (CD11cCre+/FOXO1L/L) and control mice (CD11cCre-/FOXO1L/L ) to determine whether FOXO1 deletion in DCs modulates the host response to P. gingivalis, osteoclastogenesis and periodontal disease progression. Experiments using the same two in vivo models will determine whether Akt is necessary to prevent a hyperinflammatory response in DC. These experiments will examine experimental (CD11cCre+/FOXO1L/L) and control (CD11cCre-/FOXO1L/L) mice. In Aim 3 the mechanisms by which the FOXO1-Akt axis regulates selected target gene expression (IL-1, TNF, IL-6, and IL-12) will be examined in vitro.

描述（由申请人提供）：树突状细胞（DC）通过参与将细菌刺激与牙周骨质流失联系起来的事件，在牙周疾病的发病机理中起核心作用。他们可以通过生产调节适应性和先天免疫反应的因素来实现这一目标。此外，据报道，未成熟的DC充当破骨细胞前体。在体外检查DCS的初步数据表明，转录因子FOXO1对于LPS上调了几种细胞因子IL-1，TNF，IL-6和IL-12和抗炎细胞因子IL-10的下调至关重要。此外，AKT通过减少FOXO1核定位和限制DC中的炎症性细胞因子产生，在此过程中起着重要作用。因此，我们建议FOXO1对于上调DC中的炎症反应至关重要，并且AKT阻止了过度炎症反应。这些发现是当前建议的基础，在该建议中我们将在体内检验以下假设：FOXO1-AKT轴调节DC细胞因子的表达。此外，我们将确定这种调节途径是否对于刺激对牙周病原体，牙龈疟原虫的适应性免疫反应以及它是否在细菌诱导的骨质流失中起重要作用至关重要。要删除DC中的FOXO1，我们将使用Cre-Lox方法。我们已经用在CD11C启动子控制下表达CRE重组酶的小鼠饲养了Flox的FoxO1小鼠。与同窝对照小鼠（CD11CCRE-/F​​OXO1L/L）相比，从所得的实验（CD11CCRE+/FOXO1L/L）小鼠中分离出的DC表现出降低的LPS的细胞因子表达。我们将通过在钙典写的模型中注入牙龈疟原虫来检查体内的宿主 - 细菌相互作用。由于小鼠对牙龈疟原虫很幼稚，因此当不存在适应性免疫反应时，我们可以检查反应，并将其与小鼠进行比较，在这种小鼠中，通过与牙龈疟原虫的预免疫激活了自适应免疫反应。通过流式细胞仪，我们将对FOXO1缺失在产生系统性和局部适应性免疫反应中的影响进行详细分析。还将在组织学上检查潜在的颅骨，以研究随后对破骨细胞生成和骨吸收的影响。在实验（CD1CCRE+/FOXO1L/L）和对照小鼠（CD1CCRE-/F​​OXO1L/L）中，将研究将牙龈疟原虫的口服饲料模型研究到口腔中，以确定DCS中的FOXO1缺失是否会调节对宿主对Gingivalis P. gingivalis，osteoclastasenise和perecernalsalisalise Cropersys的宿主反应。使用相同两个体内模型的实验将确定是否需要AKT来防止DC中的高炎反应。这些实验将检查实验（CD11CCRE+/FOXO1L/L）和对照（CD11ccre-/foxo1l/L）小鼠。在AIM 3中，将在体外检查FOXO1-AKT轴调节选定的靶基因表达（IL-1，TNF，IL-6和IL-12）的机制。