Oral Colonization of Aggregatibacter in Primates

描述（由申请人提供）：伴放线菌聚集杆菌（Aa）与年轻成人的局部侵袭性牙周炎（ORG）密切相关。此外，Aa具有多种毒力性状，这些毒力性状与发生在大肠杆菌中的致病事件一致。我们的小组一直在研究与Aa诱导的疾病启动相关的基因，并因此编码负责Aa附着，定植，持久性和龈下存活的毒力性状。到目前为止，我们已经归因于两个Aa自身转运蛋白粘附素基因（aae和apiA）的功能，这与Aa附着到上皮细胞的特异性有关，并表明这些粘附素（以及白细胞毒素）对旧世界（OW）灵长类动物和人类显示出物种特异性。这种特异性使得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，并着眼于预防策略的发展。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）的口腔定殖和持久性。在引入猴口腔后的28天内，将在不同时间检查每种菌株的模式、附着水平和定殖。目的1将描述的地形和定量水平的Aa发现在口腔的Rh猴最初，然后比较定植和持久性的两种菌株的Aa（一个人和一只猴子）。在放置之前，猴子将接受刮治和预防加洗必泰治疗。动物将以薄饼形式饲喂Aa，并在饲喂后28天分析定殖和持久性。选择在牙齿上定殖并持续存在的Aa，其水平等于或大于1 x 102 /mL Aa菌落，用于目标2。目的2将评估在BEC、舌和牙齿上发现的Aa的位置、水平和时间，比较未接种和野生型接种的Aa与具有aae和apiA突变的Aa，aae/apiA、flp（纤维状外蛋白）和ltx的双敲除，超过28天。这些实验应该揭示这些基因的重要性，在有关的Aa附件BEC（aae和apiA），牙齿定植（flp）和龈下生存（ltx）在口腔中的OW灵长类动物。建立该模型的效用，使我们不仅可以解剖出附着因子，但也解开其他Aa基因的免疫调节因子，因为它们会影响Aa的发病机制，在未来着眼于预防策略。

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.