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）：牙周疾病（PD）是一种常见的口腔状况，是牙齿脱落的主要原因。这是一种多数菌感染，其特征是细菌定植刺激的牙龈炎症，导致软组织破坏和肺泡 骨吸收。有趣的是，单纯疱疹病毒1（HSV-1）经常与牙周病变有关。人们认为这是一个联合因素，可以通过损害牙龈组织并维持局部炎症来增加个人对PD的敏感性。已知唾液中含有抗病毒和抗菌剂，包括作用于HSV病毒体以防止感染的成分。我们第一次观察到来自某些个体而不是其他人的唾液包含一个成分，该成分增加了牙龈成纤维细胞对HSV-1感染的敏感性。这种作用可以有利于病毒扩散并扩大对牙周病变中暴露的细胞的HSV向流。对新型唾液作用的这种意外观察促使我们在此应用程序中进一步探讨了它。因此，这项研究的目标是确定该因素的影响，以了解HSV-1如何有效地在口腔中传播，以及如何加剧PD的进展。提出了两个目标来实现这些目标：在AIM 1中，我们将确定增加细胞感染易感性的唾液蛋白。我们将结合生化分馏和功能感染测定法，以净化活性剂。鉴定将通过质谱，2D-凝胶电泳和活性纯化的馏分进行生化测定。新的初步数据表明，jacalin耗尽了唾液 它的活性表明碳水化合物在糖蛋白，粘蛋白和/或IgA上的作用。因此，我们将检查碳水化合物的作用，并使用该凝集素来改善纯化。我们还将通过研究分泌IgA的TH作用来确定HSV事先感染是否与唾液的刺激作用有关。 在AIM 2中，我们将首先定义基因表达如何受到口服各种细胞中唾液的影响。我们已邀请专家协助分析微阵列数据。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下刺激HSV-1进入。最后，为了将这种作用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。我们可以通过宾夕法尼亚州的合作者来访问各种此类细胞和病毒。 作用于口腔细胞以有利于HSV-1感染的唾液剂的鉴定为新的先例。它将开放新的调查领域，并促进研究HSV-1在PD发展中的作用。由于所有个人的唾液中都不存在该活动，因此我们将能够将其作为生物标志物进一步测试，以易于口服疱疹和可能的PD。该药物将为治疗干预措施提供一个可访问的目标，以防止HSV在口腔中扩散并限制其潜在的牙周炎活性。 公共卫生相关性：牙周疾病（PD）是一种由炎症和口腔细菌的增殖引起的多种疾病，影响了约30％的美国成年人。单纯疱疹病毒1（HSV-1）与PD病变有关，被认为是疾病发展的辅助因子。通过确定增加口腔细胞对HSV-1感染的敏感性并表征细胞对唾液的敏感性的唾液成分，本研究将：1）确定可能的生物标志物，以使口服疱疹的敏感性； 2）发现新目标 用于HSV-1感染和可能的牙周疾病的新型治疗方法。