UKY DENTAL COBRE: ORAL INFECTIONS: VIRAL INFECTIONS IN CHRONIC ORAL DISEASES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 虽然细菌是牙周炎的主要病原体，但人类疱疹病毒与炎症的牙周组织有据称的关系。这一点可能很重要，因为疱疹病毒通过各种机制调节宿主的免疫反应。这项研究的目的是验证疱疹病毒增强牙周病发病机制的假设。我们的假设正在用三种方法进行检验：1)通过分析患有和不患有牙周病的患者的临床样本，确定疱疹病毒在牙周炎中的作用；2)在大鼠模型中研究疱疹病毒促进牙周炎进展的能力；以及3)确定巨细胞病毒(CMV)的免疫调节机制。为了解决第一部分，我们正在分析健康和病变部位的牙菌斑以及100名牙周病患者的平行唾液样本。实时荧光定量聚合酶链式反应用于检测6个月以上纵向采集的样本中的人类疱疹病毒DNA。在第二个项目中，获得了大鼠巨细胞病毒(RCMV)马斯特里赫特株，并对其进行了繁殖和滴定。动物已经接种了牙龈假单胞菌和/或RCMV，以建立感染并评估牙周反应。在研究CMV免疫调节的分子机制方面，我们克隆并鉴定了CMV vIL-10基因的新亚型，最近又克隆了EBV vIL-10基因。由于这些病毒可以调节免疫反应，我们已经开始了研究，以确定它们在调节口腔上皮和牙龈成纤维细胞对牙周细菌病原体入侵的宿主反应中的作用。以人原代巨噬细胞和THP-1来源的巨噬细胞为模型，研究了CMV和EBV感染对4种口腔微生物：牙龈假单胞菌(Pg)和伴生放线菌(AA)、条件致病菌核杆菌(Fn)和非致病菌戈登葡萄球菌(Sg)对巨噬细胞功能的影响，并证实了CMV和EBV改变了巨噬细胞对这些口腔微生物的TNFa应答。我们正在确定巨细胞病毒和/或EB病毒的裂解基因是否需要改变的反应，以及这些病毒是否也抑制巨噬细胞对这些细菌的吞噬活性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Although bacteria are the primary pathogens in periodontitis, human herpesviruses have a purported relationship with inflamed periodontal tissues. This could be important because herpesviruses modulate host immune responses through a variety of mechanisms. The goal of this research was to test the hypothesis that herpesviruses enhance the pathogenesis of periodontal diseases. Our hypothesis is being examined using three approaches: 1) determine the role of herpesviruses in periodontitis by analyzing clinical samples from patients with and without periodontal disease, 2) investigating the ability of herpesvirus to enhance the progression of periodontitis in the rat model; and 3) define mechanisms of immunomodulation by cytomegalovirus (CMV). To address component 1, we are analyzing dental plaque from healthy and diseased sites and parallel saliva samples from 100 patients with periodontal disease. Real-time PCR is being used to detect DNA of human herpesvirus in the samples obtained longitudinally over 6 months. In the second project, the rat CMV (RCMV) Maastricht strain has been obtained, propagated and titered. Animals have been inoculated with P. gingivalis and/or RCMV to establish infections and evaluate the periodontal response. In the study of molecular mechanisms of immunomodulation by CMV, we have cloned and characterized novel isoforms of CMV vIL-10 gene and recently cloned EBV vIL-10. Because these viruses can modulate the immune response, we have initiated studies that determine their role in modulating the host response of oral epithelia and gingival fibroblasts against invasion by periodontal bacterial pathogens. Using human primary macrophages and THP-1 derived macrophage as models, we have studied the effects of CMV and EBV infection on the function of macrophage in response to four oral microorganisms: P. gingivalis (Pg) and A. actinomycetemcomitans (Aa); opportunistic pathogen, F. nucleatum (Fn); and non-pathogen S. gordonii (Sg), and have demonstrated that CMV and EBV alters the TNFa response of macrophages to these oral microorganisms. We are in the process of determining if lytic genes of CMV and/or EBV are required for the altered response and whether these viruses also inhibit the phagocytosis activity of macrophages to these bacteria.