Effects of saliva on herpes simplex virus infection of oralcells

唾液对口腔细胞单纯疱疹病毒感染的影响

基本信息

批准号：
8300388
负责人：
Claude F Krummenacher
金额：
$ 24万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-15 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8300388
关键词：
Address Adult Affect Afferent Neurons American Anti-Bacterial Agents Antiviral Agents Area Biochemical Bioinformatics Biological Assay Biological Markers Bone Resorption Bypass Carbohydrates Cell Line Cells Clinical Communicable Diseases Complex Mixtures Data Data Analyses Development Disease Epithelial Attachment Epithelial Cells Epithelium Etiology Exhibits Exposure to Fibroblasts Fractionation Gene Chips Gene Expression Genes Gingiva Glycoproteins Goals Gram-Negative Anaerobic Bacteria Grant Herpesviridae Herpesvirus 1 Human Immunity Immunoglobulin A Individual Infection Infection prevention Inflammation Inflammatory Inflammatory Infiltrate Investigation Kinetics Lectin Lesion Link Liquid substance Low Birth Weight Infant Mass Spectrum Analysis Membrane Fusion Modeling Mucins Nature Newborn Infant Oral Oral cavity Pathway interactions Patients Periodontal Diseases Periodontitis Play Predisposition Pregnant Women Proteins Reagent Recurrence Role Saliva Salivary Salivary Proteins Sampling Secretory Immunoglobulin A Severities Signal Transduction Simplexvirus Specificity Structure of trigeminal ganglion Surveys Testing Therapeutic Intervention Time Tissues Tooth Loss Tooth structure Tropism Two-Dimensional Gel Electrophoresis Viral Virion Virus Virus Diseases alveolar bone antimicrobial drug cardiovascular disorder risk cell injury cofactor cytokine improved in vivo jacalin latent infection microbial novel novel therapeutic intervention oral bacteria oral condition oral infection particle prevent receptor response soft tissue virology

项目摘要

DESCRIPTION (provided by applicant): Periodontal disease (PD) is a common oral condition and a leading cause of tooth loss. It is a polymicrobial infection characterized by gingival inflammation stimulated by bacterial colonization resulting in soft tissue destruction and alveolar bone resorption. Interestingly, herpes simplex virus 1 (HSV-1) is frequently associated with periodontal lesions. It is thought to be a co-factor that may increase an individual's susceptibiliy to PD by damaging gingival tissue and sustaining local inflammation. Saliva is known to contain antiviral and antimicrobial agents, including components that act on the HSV virion to prevent infection. For the first time, we observed that saliva from some individuals but not others contains a component that increases the susceptibility of gingival fibroblasts to HSV-1 infection. This effect can favor viral spread and broaden HSV tropism to cells that are exposed in periodontal lesions. This unexpected observation of a novel salivary effect prompted us to explore it further in this application. Thus, the goals of this study are to identify this factor ad its effects in order to understand how HSV-1 efficiently spreads in the oral cavity and how it may exacerbate the progression of PD. Two aims are proposed to achieve these objectives: In aim 1, we will identify the salivary protein that increases cell susceptibility to infection. We will combine biochemical fractionation and functional infection assays to purify the active agent. Identification will be performed by mass spectrometry, 2D-gel electrophoresis and biochemical assays on active purified fractions. Novel preliminary data showing that jacalin depletes saliva of its activity suggest a role for carbohydrates on glycoproteins, mucins and/or IgA. Thus we will examine the role of carbohydrates and use this lectin to improve purification. We will also determine if prior infection by HSV relates to the stimulatory effect of saliva by investigating th role of secretory IgA. In aim 2, we will first define how gene expression is affected by exposure to saliva in various cells of oral origin. We have enlisted experts to aid in analyzing microarray data. We will also use model cell lines to define how saliva stimulates HSV-1 entry at the level of receptor interaction, entry pathways and membrane fusion. Finally, to relate this effect to in vivo infection, we will survey primary human oral cells and HSV clinical isolates. We have access to a variety of such cells and viruses by virtue of collaborators here at PENN. The identification of a salivary agent that acts on oral cells to favor HSV-1 infection sets a new precedent. It will open new areas of investigation and facilitate studies of the role of HSV-1 in the progression of PD. Since the activity is not present in the saliva from all individuals, we wil be able to further test it as a biomarker for susceptibility to oral herpes and possibly PD. This agent will provide an accessible target for therapeutic interventions to prevent HSV spread in the oral cavity and limit its potential aggravating activity in periodontitis. PUBLIC HEALTH RELEVANCE: Periodontal disease (PD) is a polymicrobial disease caused by inflammation and proliferation of oral bacteria that affects around 30% of American adults. Herpes simplex virus 1 (HSV-1) is associated with PD lesions and is thought to be a cofactor in the development of the disease. By identifying salivary components that increase susceptibility of oral cells to HSV-1 infection and by characterizing the cellular response to saliva, this study will: 1) identify possible biomarkers for susceptibility to oral herpes; and 2) uncover new targets for novel therapeutic approaches to HSV-1 infection and possibly periodontal diseases.

描述（由申请人提供）：牙周病（PD）是一种常见的口腔疾病，也是牙齿脱落的主要原因。它是一种多种微生物感染，其特征是细菌定植刺激牙龈炎症，导致软组织破坏和牙槽骨损伤。骨吸收。有趣的是，单纯疱疹病毒 1 (HSV-1) 经常与牙周病变相关。它被认为是一种辅助因子，可能会通过损害牙龈组织和维持局部炎症来增加个体对 PD 的易感性。众所周知，唾液含有抗病毒和抗菌剂，包括作用于 HSV 病毒颗粒以预防感染的成分。我们首次观察到某些个体（而非其他个体）的唾液中含有一种会增加牙龈成纤维细胞对 HSV-1 感染易感性的成分。这种效应有利于病毒传播并扩大HSV对牙周病变中暴露的细胞的趋向性。这种对新型唾液效应的意外观察促使我们在本应用中进一步探索它。因此，本研究的目标是确定这一因素及其影响，以便了解 HSV-1 如何在口腔中有效传播以及它如何加剧 PD 的进展。为了实现这些目标，提出了两个目标：在目标 1 中，我们将鉴定增加细胞感染易感性的唾液蛋白。我们将结合生化分离和功能感染测定来纯化活性剂。将通过质谱、2D 凝胶电泳和生化分析对活性纯化组分进行鉴定。新的初步数据显示 Jacalin 会消耗唾液其活性表明碳水化合物对糖蛋白、粘蛋白和/或 IgA 具有一定的作用。因此，我们将研究碳水化合物的作用并使用这种凝集素来改善纯化。我们还将通过研究分泌型 IgA 的作用来确定先前的 HSV 感染是否与唾液的刺激作用有关。在目标 2 中，我们将首先定义口腔来源的各种细胞中暴露于唾液如何影响基因表达。我们聘请了专家来帮助分析微阵列数据。我们还将使用模型细胞系来定义唾液如何在受体相互作用、进入途径和膜融合水平上刺激 HSV-1 进入。最后，为了将这种效应与体内感染联系起来，我们将调查原代人类口腔细胞和 HSV 临床分离株。借助宾夕法尼亚大学的合作者，我们可以获得各种此类细胞和病毒。鉴定出一种作用于口腔细胞以促进 HSV-1 感染的唾液剂开创了新的先例。它将开辟新的研究领域并促进 HSV-1 在 PD 进展中的作用的研究。由于并非所有个体的唾液中都存在这种活性，我们将能够进一步测试它作为口腔疱疹和可能的帕金森病易感性的生物标志物。该药物将为治疗干预提供一个可及的目标，以防止单纯疱疹病毒在口腔中传播并限制其潜在的牙周炎加重活动。公共卫生相关性：牙周病 (PD) 是一种由口腔细菌炎症和增殖引起的多种微生物疾病，影响约 30% 的美国成年人。单纯疱疹病毒 1 (HSV-1) 与 PD 病变相关，被认为是该疾病发展的辅助因子。通过识别增加口腔细胞对 HSV-1 感染易感性的唾液成分，并表征细胞对唾液的反应，本研究将： 1) 识别口腔疱疹易感性的可能生物标志物； 2）发现新目标针对 HSV-1 感染和可能的牙周疾病的新治疗方法。