Periodontal bacteria enhance oral KSHV pathogenesis and Kaposi's Sarcoma development in HIV + patients

项目摘要 卡波西氏肉瘤(KS)仍然是HIV/AIDS患者最常见的肿瘤，并累及 口腔是这种肿瘤最常见的临床表现之一。艾滋病毒感染会导致 增加患牙周病和各种致病菌携带口腔的风险。然而， 牙周细菌和致癌病毒在当地环境中的相互作用是否有助于 这些病毒在口腔中的复制或维持在很大程度上仍不清楚。我们出版的和 初步数据表明，人类原代口腔细胞与两种典型病原体相关 牙周主要细菌产生的分子模式(PAMP)-脂磷壁酸(LTA) 金黄色葡萄球菌(Sa)和牙龈卟啉单胞菌(Pg)脂多糖(LPS)-增加 新生KSHV感染过程中病毒的初始进入和随后的潜伏基因表达。此外，LTA和 脂多糖上调KSHV进入细胞受体之一硫酸乙酰肝素蛋白多糖(HSPG)并增加 活性氧(ROS)的产生是KSHV感染的辅助因素。另外，我们发现 来自SA和PG培养物或细菌PAMPs的条件培养液诱导病毒裂解重新激活 通过调节病毒microRNAs的表达，潜伏感染口腔细胞，促进病毒传播。为 临床意义：我们发现沙门氏菌、葡萄球菌和柯萨奇病毒在我们的口腔中有很高的感染率。 HIV+患者队列，发现唾液ROS、宿主抗氧化因子和细菌水平较高 HIV+/KSHV+患者的LTA/LPS高于HIV+/KSHV-患者。根据这些数据，我们假设 牙周细菌或其PAMP可能促进初始KSHV感染、复制和传播 在口腔中HIV+患者通过多种宿主和病毒因素，最终促进口腔 KS开发。为了解决这一假设，我们提出了以下具体目标，包括临床 和基础研究：1)确定临床相关性和宿主唾液抗氧化剂之间的相关性 HIV+患者中的因素、特定细菌携带/PAMPs水平以及KSHV口腔脱落。2)确定 牙周细菌及其PAMP促进KSHV初始感染和病毒溶解的机制 重新激活，是开发KS所必需的两个重要步骤。通过这些努力，我们可以更好地 了解病原体混合感染如何在口腔独特的利基环境中促进病毒相关癌症的发展 免疫功能低下的患者。我们的研究结果也将为临床的发展提供框架 评估干扰宿主-细菌-病毒相互作用的策略的试验(例如，特定的抗生素、靶向 TLRS-ROS轴或病毒microRNAs合成)，以减少或预防口腔KSHV感染和 HIV阳性患者的KS进展。

PROJECT ABSTRACT Kaposi's sarcoma (KS) remains the most common tumor arising in patients with HIV/AIDS, and involvement of the oral cavity represents one of the most common clinical manifestations of this tumor. HIV infection incurs an increased risk for periodontal diseases and oral carriage from a variety of pathogenic bacteria. However, whether interactions involving periodontal bacteria and oncogenic viruses in the local environment facilitate replication or maintenance of these viruses in the oral cavity remains largely unknown. Our published and preliminary data indicate that incubation of human primary oral cells with two prototypical pathogen-associated molecular patterns (PAMPs) produced by prominent periodontal bacteria—lipoteichoic acid (LTA) from Staphylococcus aureus (Sa) and lipopolysaccharide (LPS) from Porphyromonas gingivalis (Pg)—increases initial viral entry and subsequent latent gene expression during de novo KSHV infection. Moreover, LTA and LPS up-regulate one of cellular receptors for KSHV entry, Heparan sulfate proteoglycan (HSPG) and increase reactive oxygen species (ROS) production as co-factor contributed to KSHV infection. Additionally, we found that conditioned medium from Sa and Pg culture or bacterial PAMPs induced viral lytic reactivation from latently infected oral cells through regulation of viral microRNAs expression, promoting virus dissemination. For clinical relevance, we found a high infection and co-infection rate of Sa, Pg and KSHV in the oral cavity of our cohort of HIV+ patients, and found higher levels of salivary ROS, host antioxidant factors and bacterial LTA/LPS from HIV+/KSHV+ patients than those HIV+/KSHV- ones. Based on these data, we hypothesize that periodontal bacteria or their PAMPs may facilitate initial KSHV infection, replication and dissemination in the oral cavity of HIV+ patients through multiple host and viral factors, and ultimately promote oral KS development. To address this hypothesis, we propose the following specific aims including both clinical and basic studies: 1) Determine the clinical relevance and correlations between host salivary antioxidant factors, specific bacterial carriage/PAMPs levels, and KSHV oral shedding in HIV+ patients. 2) Identify the mechanisms through which periodontal bacteria or their PAMPs facilitate KSHV initial infection and viral lytic reactivation, two important steps necessary for KS development. Through these efforts, we can better understand how pathogens co-infection can promote virus-associated cancer development in oral unique niche of immunocompromised patients. Our results will also provide the framework for the development of clinical trials evaluating the strategies interfering with host-bacteria-virus interaction (e.g. specific antibiotics, targeting TLRs-ROS axis or viral microRNAs synthesis) for their abilities to reduce or prevent oral KSHV infection and KS progression in HIV+ patients.