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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 虽然细菌是牙周炎的主要病原体，但人类疱疹病毒与发炎的牙周组织有据称的关系。这可能很重要，因为疱疹病毒通过多种机制调节宿主免疫反应。 本研究的目的是验证疱疹病毒增强牙周病发病机制的假设。 我们的假设正在使用三种方法进行检验：1）通过分析牙周病患者和非牙周病患者的临床样本来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; 3）确定巨细胞病毒（CMV）的免疫调节机制。 为了解决第一部分，我们分析了健康和患病部位的牙菌斑以及100名牙周病患者的平行唾液样本。 实时荧光定量PCR方法用于检测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CMV（RCMV）马斯特里赫特株，并进行了繁殖和滴定。 用牙龈卟啉单胞菌和/或RCMV接种动物以建立感染并评价牙周反应。在CMV免疫调节的分子机制的研究中，我们已经克隆和表征了CMV vIL-10基因的新亚型，最近克隆了EBV vIL-10。由于这些病毒可以调节免疫反应，我们已经开始研究，确定它们在调节口腔上皮细胞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源性巨噬细胞为模型，研究巨噬细胞对牙龈卟啉单胞菌（Pg）和牙龈卟啉单胞菌（A. gingivalis，Pg）等4种口腔微生物的功能反应。伴放线菌（Aa）、条件致病菌F. nucleatum（Fn）和非病原S. gordonii（Sg），并且已经证明CMV和EBV改变巨噬细胞对这些口腔微生物的TNF α应答。我们正在确定CMV和/或EBV的裂解基因是否是改变反应所必需的，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.