Treponema denticola cytoskeletal filaments and oral infection

齿垢密螺旋体细胞骨架丝与口腔感染

• 批准号: 7253018
• 负责人: JACQUES G. IZARD
• 金额: $ 24.11万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7253018
• 关键词: Affect; Animal Model; Antigen Presentation; Bacteria; Bacterial Infections; Basic Science; Biology; Bone Resorption; Bone Tissue; Cell-Matrix Junction; Cells; Class; Clinical; Cytoplasmic Filaments; Cytoplasmic Structures; Cytoskeletal Filaments; Data; Development; Disease Outcome; Disease Progression; Disease model; End Point; Endodontics; Engineering; Event; Filament; Formalin; Future; Genetic Engineering; Gingivitis; Goals; Grant; Healed; Histology; Human; Immune response; In Vitro; Infection; Inflammation; Interleukin-10; Intermediate Filaments; Invasive; Knock-out; Life; Measurement; Membrane; Methodology; Microbial Biofilms; Modeling; Modification; Mouse Strains; Mus; Nature; Oral; Organism; Outcome; Pathogenesis; Pathogenicity; Patients; Penetration; Periodontal Diseases; Periodontal Infection; Periodontitis; Phase; Plant Roots; Population; Population Analysis; Population Process; Predisposition; Process; Production; Proteins; Research; Ribosomal RNA; Role; Site; Structure; Testing; Tissues; Tooth Cervix; Tooth structure; Toxin; Translational Research; Treponema; Treponema denticola; Work; alveolar bone; base; bone loss; cell motility; cell type; cytokine; experience; fighting; healing; in vivo; killings; microbial; mouse model; mutant; oral biofilm; oral infection; oral pathogen; oral spirochetes; pathogen; periplasm; pressure; prevent; research study; tool

DESCRIPTION (provided by applicant): Among the key pathogens involved in oral bacterial diseases, Treponema denticola is the most invasive organism. The various oral Treponema bacterial species have in common a cytoplasmic structure with organizational similarities to intermediate filaments. In vitro experiments have demonstrated the importance of such structure in key events related to oral infectivity. The goal of this research is to decipher the importance and function of treponemal intermediate-like filaments in pathogenicity in vivo. The long-term goal is to develop new strategies to combat the invasive nature of treponemal cells, while maintaining a healthy oral microbiota. The rationale is to validate the hypothesis that treponemes propagation (colonization, persistence, and dissemination first local and then systemic) is dependent upon intermediate- like filaments. The proposed studies will focus on testing the pathogenicity of a genetically engineered T. denticola mutant deficient in intermediate-like filaments (cfpA knockout) in appropriate animal models to study oral infections. The infectivity of the cfpA knockout mutant will be tested in two mouse models. The first mouse model investigates the tissue and bone damage associated with an endodontic infection (Aim 1). The second mouse model tests the pathogenicity in a periodontal infection (Aim 2). In conjunction with the cfpA knockout mutant, the wild-type strain (live and formalin-killed) will be used as control, as well as a mutant deficient in motility to test the effect of live antigen presentation during the colonization step. The experimental quantitative endpoint is the measurement of bone loss at the root of the infected tooth in the endodontic model, and the recession of the alveolar bone crest from the cementoenamel junction in the periodontal model. A qualitative criterion is the inflammation associated with the infection, which is to be evaluated by histology of the center plane of the infected tooth in the endodontic infection model, and by cytokine production in the periodontal infection model. In the case of the periodontal infection model, an additional criterion will be the modification of the oral microbiota post-infection. This study will further our understanding of the pathogenesis of T. denticola in both periodontal and endodontic infections by providing a more complete understanding of the role of intermediate-like filaments in infectivity, the importance of motility for treponemes persistence, and model the biofilm population changes associated with periodontal disease. To allow the modulation of the oral biofilm, clinicians need new strategies to fight periodontal diseases. New strategies rely on an understanding of microbial biology. To limit bone loss and other damages, proper host responses and healthy natural biofilm should be promoted in the patient healing process. Our strategy is to target key functional entities of aggressive and invasive pathogens, with the goal of eliminating the damage- inducing population.

描述(申请人提供)：在涉及口腔细菌性疾病的关键病原体中，齿密螺旋体是侵袭性最强的生物。各种口腔密螺旋体细菌具有共同的细胞质结构，其组织结构类似于中间丝。体外实验已经证明了这种结构在与口腔传染性相关的关键事件中的重要性。本研究的目的是破译梅毒螺旋体中间样丝在体内致病中的重要性和功能。长期目标是开发新的策略来对抗梅毒螺旋体细胞的侵袭性，同时保持健康的口腔微生物区系。其基本原理是验证这样的假设，即密螺旋体的传播(定殖化、持久性和传播首先是局部的，然后是全身性的)依赖于类似中间的细丝。拟议的研究将集中于在适当的动物模型中测试缺乏中间类细丝(CFPA基因敲除)的基因工程齿状毛滴虫突变体的致病性，以研究口腔感染。CFPA基因敲除突变体的传染性将在两个小鼠模型中进行测试。第一个小鼠模型研究与牙髓感染相关的组织和骨骼损伤(目标1)。第二个小鼠模型测试牙周感染的致病性(目标2)。与CFPA基因敲除突变体一起，野生型菌株(活的和福尔马林灭活的)将作为对照，以及一个活力不足的突变体，以测试在殖民步骤中活的抗原呈递的影响。实验的定量终点是测量牙髓模型中感染牙根部的骨丢失，以及牙周模型中牙槽骨从牙釉质交界处的后退。定性的标准是与感染相关的炎症，在牙髓感染模型中，通过感染牙齿中心平面的组织学来评价，在牙周感染模型中，通过细胞因子的产生来评价。在牙周感染模型的情况下，另一个标准将是感染后口腔微生物区系的修改。这项研究将使我们更全面地了解齿状毛滴虫在牙周和牙髓感染中的致病机制，更全面地了解中间丝在感染性中的作用，运动对密螺旋体持续存在的重要性，并模拟与牙周病相关的生物被膜群的变化。为了能够调节口腔生物膜，临床医生需要新的策略来对抗牙周病。新的策略依赖于对微生物生物学的理解。为了减少骨丢失和其他损害，在患者的愈合过程中应该促进适当的宿主反应和健康的自然生物膜。我们的战略是以侵袭性和侵袭性病原体的关键功能实体为目标，目标是消除导致损害的种群。