Host Response to Challenge with Porphyromonas gingivalis DNA

描述（由申请人提供）：牙周病是最常见的人类疾病之一，困扰着15%的人口。最近的数据表明牙周炎是许多其他系统性并发症的病因。牙龈卟啉单胞菌被认为是最强的牙周致病菌之一，也是牙周病进展的病原体。在没有疾病的情况下，它也存在于口腔中。虽然细菌与宿主的相互作用对于理解疾病机制非常重要，但对参与相互作用的宿主和病原体组分仍然知之甚少。体外研究表明，牙龈卟啉单胞菌可以侵入多种细胞类型，包括牙龈上皮细胞。它还被证明可以防止这些细胞的凋亡，复制和传播到周围的细胞。有趣的是，牙龈卟啉单胞菌已被证明在健康和易感宿主中进行这些活动。研究还揭示了牙龈卟啉单胞菌的复杂性质，这是由于根据宿主细胞类型、细菌菌株、初始接种和使用活（代谢活性）与死细菌或特定细菌组分（例如半胱氨酸蛋白酶，LPS）的存在而产生的不同结果以及与之密切相关。几项研究集中在Toll样受体介导的宿主细胞应答上，这些受体被细菌组分激发，以了解先天免疫应答的启动。在这些细菌成分中，很少有人关注DNA，特别是口腔上皮细胞。为了进一步了解牙龈卟啉单胞菌的发病机制，我们将重点关注细菌的一种成分，DNA。本研究的具体目的包括：（1）表征两种口腔上皮细胞系和从两名患者收集的原代细胞对牙龈卟啉单胞菌W83 DNA攻击的反应，（2）TLR9在介导这种反应中的作用，以及（3）可能在这种相互作用中发挥作用的其他DNA受体。初步的体外研究表明，口腔上皮细胞系HN 4对来自牙龈卟啉单胞菌的DNA的攻击有应答，并在转录水平上进行基因调控。此外，我们已经证明了HN 4细胞摄取CpG寡核苷酸。一项额外的初步研究表明，在用牙龈卟啉单胞菌DNA攻击的TLR9敲低的HN 4细胞系中，基因调控似乎没有受到显著影响，因此，我们计划检查胞质DNA受体DAI，其也可能在牙龈卟啉单胞菌的发病机制中发挥作用。 公共卫生相关性：这项研究将导致更好地了解宿主反应的机制，有助于牙龈卟啉单胞菌W83的发病机制。这些研究可能会揭示预防牙周病的潜在目标。

DESCRIPTION (provided by applicant): Periodontal disease is one of the most common human diseases afflicting 15% of the population. Recent data has implicated periodontitis as a causative disease for many other systemic complications. Porphoromonas gingivalis is named one of the most virulent periodontopathogens and an etiological agent in the progression of this disease. It also is present in the oral cavity in the absence of the disease. Although the interaction of the bacterium with the host is of major importance for the understanding of the disease mechanisms, both the host as well as the pathogen components involved in the interaction remain poorly understood. In vitro studies have revealed that P. gingivalis can invade a variety of cell types including gingival epithelial cells. It has also been demonstrated to prevent apoptosis in these cells, replicate, and disseminate to surrounding cells. Interestingly, P. gingivalis has been shown to perform these activities in both healthy and susceptible hosts. Studies have also reveled the complex nature of P. gingivalis due to varying results according to, and closely associated with; the host cell type, bacterial strain, initial inoculation, and the use of live (metabolically active) versus dead bacteria, or the presence of specific bacterial components (e.g. cysteine proteinases, LPS). Several studies have focused on the host cell responses mediated by Toll-like receptors challenged with bacterial components to gain insight on the initiation of the innate immune response. Of these bacterial components, little attention has been paid to DNA, especially in oral epithelial cells. To further understand the pathogenesis of P. gingivalis, we will be focusing on one bacterial component, DNA. The specific aims of this study include: (1) The characterization of the response of two oral epithelial cell lines and primary cells harvested from two patients to challenge with P. gingivalis W83 DNA, (2) The role of TLR9 in mediating this response, and (3) other DNA receptors that may play a role in this interaction. Initial in vitro studies revealed that the oral epithelial cell line HN4 responds to challenge with DNA derived from P. gingivalis with gene regulation at the transcriptional level. In addition, we have demonstrated that HN4 cells uptake CpG oligonucleotides. An additional preliminary study has shown that gene regulation does not seem to be significantly impacted in TLR9 knockdown HN4 cell lines that were challenged with P. gingivalis DNA, therefore, we plan to examine the cytosolic DNA receptor DAI, which also may play a role in the pathogenesis of P. gingivalis. Public health Relevance: This study will lead to a greater understanding of the mechanisms of the host response that contribute to the pathogenesis of P. gingivalis W83. These studies may then reveal potential targets for preventing the initiation of periodontal disease.