Oral Colonization of Aggregatibacter in Primates

灵长类动物中聚集杆菌的口腔定植

• 批准号: 8249833
• 负责人: DANIEL H FINE
• 金额: $ 4.21万
• 依托单位: RBHS-SCHOOL OF DENTAL MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8249833
• 关键词: Actinobacillus actinomycetemcomitans; Address; Affect; Animals; Antibiotics; Bacteria; Bacterial Adhesins; Binding; Cells; Child; Chlorhexidine; Data; Debridement; Development; Disease; Environment; Epithelium; Event; Eye; Future; Gene Proteins; Genes; Genetic; Goals; Host Defense; Hour; Human; Infection; Infection prevention; Infectious Skin Diseases; Intercept; Knock-out; Label; Location; Macaca mulatta; Modeling; Modification; Monkeys; Movement; Mutation; Oral; Oral cavity; Pathogenesis; Pattern; Periodontitis; Play; Positioning Attribute; Prevention strategy; Primates; Prophylactic treatment; Proteins; Research; Research Design; Resort; Role; Sampling; Serotyping; Site; Spatial Distribution; Species Specificity; Specificity; Spectinomycin; Staging; Study models; Surface; Testing; Time; Tissues; Tongue; Tooth Loss; Tooth structure; Virulence; base; design; feeding; immunoregulation; killings; leukotoxin; man; migration; premature; prevent; public health relevance; research study; trait; young adult

DESCRIPTION (provided by applicant): Aggregatibacter actinomycetemcomitans (Aa) is intimately associated with Localized Aggressive Periodontitis (LAP) in young adults. Further, Aa possesses a variety of virulence traits that are consistent with pathogenic events that occur in LAP. Our group has been studying genes that are related to Aa-induced disease initiation and as such encode virulence traits responsible for Aa attachment, colonization, persistence, and subgingival survival. Thus far we have ascribed functions for two Aa autotransporter adhesin genes (aae and apiA) that are related to the specificity of Aa attachment to epithelium and have shown that these adhesins (as well as Leukotoxin) show species specificity for Old World (OW) primates and humans. This specificity makes the OW primate an ideal model for studying early events related to Aa infection with an eye toward development of preventive strategies. This R21 application consists of two Aims and is designed to compare oral colonization and persistence of two Aa strains (one from humans and one from Rhesus [Rh] monkeys) that are introduced into the oral cavity of OW primates. Each strain will be examined for its pattern, level of attachment, and colonization at different times over a 28-day period following introduction into the mouth of the monkey. Aim 1 will describe the topographical and quantitative level of Aa found in the mouths of Rh monkeys initially and then compare colonization and persistence of the two strains of Aa (one human and one monkey). Before placement, monkeys will receive scaling and prophylaxis plus chlorhexidine treatment. Animals will be fed Aa in a pancake and colonization and persistence will be analyzed 28 days after feeding. The Aa that colonizes and persists on teeth at a level equal to or greater than 1 x 102 /mL colonies of Aa will be selected for use in Aim 2. Aim 2 will assess the location, level and timing of Aa found on BECs, tongue and teeth comparing un-inoculated and wild type inoculated Aa to Aa with mutations in aae and apiA, a double knockout of aae/apiA, flp (the fibrillar outer protein) and ltx over a 28-day period. These experiments should reveal the importance of these genes in relation to Aa attachment to BECs (aae and apiA), to tooth colonization (flp) and to subgingival survival (ltx) in the oral cavity of OW primates. Establishing the utility of this model should allow us not only to dissect out attachment factors but also to unravel immune modulation factors from other Aa genes as they affect Aa pathogenesis in the future with an eye toward preventive strategies. PUBLIC HEALTH RELEVANCE: Aggregatibacter actinomycetemcomitans is intimately associated with Localized Aggressive Periodontitis in young children, which can result in premature loss of teeth. Coincidentally, A. actinomycetemcomitans is also found in the mouths of Old World primates such as Rhesus (Rh) monkeys. It has been shown that A. actinomycetemcomitans attaches to tissues and kills defense cells in humans and Rh monkeys in a very similar manner. Therefore the Rh monkey provides us with an ideal model in our efforts to unravel some of the mysteries related to the earliest stages of A. actinomycetemcomitans-induced disease. It is also known that A. actinomycetemcomitans produces several factors that are similar to many other bacteria that cause severe infections in man. Therefore, studying A. actinomycetemcomitans in the mouth of Rh monkeys can provide a model that can help us understand how A. actinomycetemcomitans-induced infections develop in a real world environment. This understanding may also have application for other mucosal diseases. We have developed a relationship with the Northeast Primate Research Center and have designed a study that will enable us to place this bacterium in the mouths of Rh monkeys. This study design will allow us to examine how genes direct the way in which A. actinomycetemcomitans attaches to tissues and initiates disease. While attachment is recognized as the first step in infection of skin surfaces that include surfaces like the gums, survival depends on the ability of the bacteria to defend itself against host defense cells. This proposal is designed to examine how certain A. actinomycetemcomitans genes influence early events such as attachment and survival below the gum line. Use of this primate model will allow us to develop ways to interfere with attachment and survival in the hope that we can devise strategies to prevent infections without the need to resort to the use of antibiotics. )

描述（由申请人提供）：聚集的放线菌Cetemcitans（AA）与年轻人的局部攻击性牙周炎（LAP）密切相关。此外，AA具有多种与膝上发生的致病事件一致的毒力性状。我们的小组一直在研究与AA诱导的疾病起始有关的基因，并因此编码了负责AA附着，定殖，持久性和亚生生存的毒力特征。到目前为止，我们已经将两个AA自转移蛋白基因（AAE和APIA）归因于与AA附着在上皮细胞上的特异性有关的功能，并表明这些粘附素（以及白细胞毒素）表现出针对旧世界（OW）灵长类动物和人类的物种特异性。这种特异性使OW灵长类动物成为研究与AA感染相关的早期事件的理想模型，并着眼于开发预防策略。该R21应用程序由两个目标组成，旨在比较两个AA菌株的口腔定植和持久性（一种来自人类，另一个来自恒河猴[Rh]猴子），这些菌株旨在引入OW灵长类动物的口腔腔。在引入猴子口之后的28天期间，将检查每个菌株的模式，依恋水平和殖民水平和定殖。 AIM 1将描述最初在RH猴子口中发现的AA的地形和定量水平，然后比较AA的两种菌株的殖民化和持久性（一个人和一只猴子）。放置之前，猴子将接受缩放和预防和氯己定治疗。动物将在煎饼中喂食AA，并在喂食后28天分析殖民化和持久性。将在AIM 2中选择与AA的牙齿上定居并持续在牙齿上的AA。 LTX在28天内。这些实验应揭示这些基因与AA与BEC（AAE和APIA），牙齿定殖（FLP）的重要性以及在OW灵长类动物的口腔腔中的尺寸生存（LTX）。建立该模型的实用性不仅应使我们不仅可以剖析附着因素，而且还可以从其他AA基因中解散免疫调节因子，因为它们将来会影响AA发病机理，并着眼于预防策略。 公共卫生相关性：聚集的放线菌ctemcetemcitans与幼儿的局部侵袭性牙周炎密切相关，这可能导致牙齿过早损失。巧合的是，在恒河猴（RH）猴子等旧世界灵长类动物的口中也发现了A.放线菌。已经表明，曲霉的放线菌与组织相连，并以非常相似的方式杀死了人类和RH猴子的防御细胞。因此，RH猴子为我们提供了一个理想的模型，以揭示与曲霉曲霉诱发的疾病最早阶段有关的一些奥秘。众所周知，曲霉曲霉的人会产生与许多其他细菌相似的因素，这些因素会引起人类严重感染。因此，在RH猴口研究A.放线菌ceTEMCOMITAN可以提供一个模型，可以帮助我们了解A. actinomycetemcomitans诱导的感染如何在现实世界环境中发展。这种理解也可能应用于其他粘膜疾病。我们已经与东北灵长类动物研究中心建立了关系，并设计了一项研究，该研究将使我们能够将这种细菌置于RH猴子的口中。这项研究设计将使我们能够研究基因如何指导A.放线菌cotemcetemcomitans对组织的附着并造成疾病的方式。虽然依恋被认为是包括牙龈等表面感染的第一步，但生存取决于细菌防御自身防御宿主防御细胞的能力。该提案旨在研究某些A.放线菌的基因如何影响早期事件，例如牙龈线以下的附着和生存。使用这种灵长类动物模型将使我们能够开发出干扰依恋和生存的方法，希望我们可以制定策略以防止感染而无需诉诸使用抗生素。 ）