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

• 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
• 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

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 骨吸收有趣的是，单纯疱疹病毒1型（HSV-1）经常与牙周病变有关。它被认为是一种辅助因子，可能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。已知唾液含有抗病毒剂和抗微生物剂，包括作用于HSV病毒体以防止感染的组分。这是第一次，我们观察到，唾液从一些人，而不是其他人含有一种成分，增加牙龈成纤维细胞对HSV-1感染的易感性。这种效应可以促进病毒传播，并扩大HSV对牙周病变中暴露的细胞的嗜性。这一新的唾液效应的意外观察促使我们在本应用中进一步探索。因此，本研究的目的是确定这一因素及其影响，以了解HSV-1如何有效地在口腔中传播，以及它如何加剧PD的进展。提出了两个目标来实现这些目标：在目标1中，我们将确定唾液蛋白，增加细胞对感染的易感性。我们将结合联合收割机生化分离和功能感染试验来纯化活性剂。将通过质谱法、2D凝胶电泳和生化试验对活性纯化组分进行鉴别。新的初步数据显示，jacalin消耗唾液的 其活性表明碳水化合物对糖蛋白、粘蛋白和/或伊加作用。因此，我们将研究碳水化合物的作用，并使用这种凝集素，以提高纯化。我们还将通过研究分泌型伊加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，我们将首先定义基因表达如何受到口腔来源的各种细胞中唾液暴露的影响。我们已经招募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、进入途径和膜融合的水平上刺激HSV-1进入。最后，为了将这种效应与体内感染联系起来，我们将调查原代人口腔细胞和HSV临床分离株。我们有机会获得各种这样的细胞和病毒凭借合作者在这里在宾夕法尼亚州。 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一个新的先例。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。由于活性并不存在于所有个体的唾液中，我们将能够进一步测试它作为口腔疱疹易感性和可能的PD的生物标志物。该制剂将为预防HSV在口腔中的传播和限制其在牙周炎中的潜在加重活性的治疗干预提供可接近的靶点。 公共卫生相关性：牙周病（PD）是一种由口腔细菌的炎症和增殖引起的多微生物疾病，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，被认为是疾病发展的辅助因素。通过鉴定增加口腔细胞对HSV-1感染易感性的唾液成分，并通过表征对唾液的细胞反应，本研究将：1）鉴定口腔疱疹易感性的可能生物标志物; 2）发现新的靶点 用于治疗HSV-1感染和可能的牙周疾病的新方法。