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）：放线菌聚集杆菌 (Aa) 与年轻人的局限性侵袭性牙周炎 (LAP) 密切相关。此外，Aa 具有多种与 LAP 中发生的致病事件一致的毒力特征。我们的小组一直在研究与 Aa 诱导的疾病发生相关的基因，这些基因编码负责 Aa 附着、定植、持久性和龈下存活的毒力特征。到目前为止，我们已经确定了两个 Aa 自转运粘附素基因（aae 和 apiA）的功能，它们与 Aa 附着到上皮的特异性相关，并表明这些粘附素（以及白细胞毒素）对旧世界 (OW) 灵长类动物表现出物种特异性和人类。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并着眼于制定预防策略的理想模型。该 R21 应用程序由两个目标组成，旨在比较引入 OW 灵长类动物口腔的两种 Aa 菌株（一种来自人类，一种来自恒河猴 [Rh] 猴）的口腔定植和持久性。每种菌株在引入猴子口腔后的 28 天内，将在不同时间检查其模式、附着水平和定植情况。目标 1 将首先描述在 Rh 猴口腔中发现的 Aa 的分布和定量水平，然后比较两种 Aa 菌株（一只人类和一只猴子）的定植和持久性。在安置之前，猴子将接受刮治和预防以及氯己定治疗。动物将被喂食煎饼中的 Aa，并在喂食后 28 天分析定植和持久性。将选择在牙齿上定殖并持续存在水平等于或大于 1 x 102 /mL Aa 菌落水平的 Aa 用于目标 2。目标 2 将评估 BEC、舌头上发现的 Aa 的位置、水平和时间和牙齿，比较未接种和野生型接种的 Aa 与 aae 和 apiA 突变的 Aa，aae/apiA、flp（纤维状外部蛋白）的双敲除和 28 天内的 ltx。这些实验应该揭示这些基因对于 OW 灵长类动物口腔中 Aa 与 BEC（aae 和 apiA）的附着、牙齿定植 (flp) 和龈下存活 (ltx) 的重要性。建立该模型的实用性不仅可以让我们剖析附着因子，还可以解开其他 Aa 基因的免疫调节因子，因为它们将来会影响 Aa 发病机制，并着眼于预防策略。 公众健康相关性：放线菌聚集菌与幼儿局部侵袭性牙周炎密切相关，可导致牙齿过早脱落。巧合的是，A. actinomycetemcomitans 也存在于旧大陆灵长类动物（例如恒河猴）的口腔中。研究表明，A. actinomycetemcomitans 以非常相似的方式附着在组织上并杀死人类和 Rh 猴的防御细胞。因此，Rh 猴为我们提供了一个理想的模型，以帮助我们解开与 A. actinomycetemcomitans 引起的疾病的最早阶段相关的一些谜团。还已知，伴随放线放线菌产生的多种因子与许多其他导致人类严重感染的细菌相似。因此，研究Rh猴口腔中的A. actinomycetemcomitans可以提供一个模型，帮助我们了解A. actinomycetemcomitans诱导的感染如何在现实世界环境中发生。这种理解也可能适用于其他粘膜疾病。我们与东北灵长类动物研究中心建立了合作关系，并设计了一项研究，使我们能够将这种细菌放入 Rh 猴的嘴里。这项研究设计将使我们能够检查基因如何指导放线放线菌附着在组织上并引发疾病的方式。虽然附着被认为是皮肤表面（包括牙龈等表面）感染的第一步，但细菌的生存取决于细菌抵御宿主防御细胞的能力。该提案旨在研究某些放线放线菌基因如何影响早期事件，例如牙龈线以下的附着和存活。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，希望我们能够设计出预防感染的策略，而无需诉诸抗生素。 ）