Study of Human Bocavirus Infection in Human Airway Epithelia

人气道上皮细胞博卡病毒感染的研究

• 批准号: 8638274
• 负责人: Jianming Qiu
• 金额: $ 20.15万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-18 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8638274
• 关键词: 2 year old; Acute; Adenoviruses; Antiviral Agents; Apical; Apoptosis; Cell Death; Cells; Characteristics; Child; Childhood; Clinical; Collaborations; DNA Damage; DNA Viruses; Detection; Developed Countries; Development; Embryo; Epithelial; Epithelial Cells; Event; Family; Foundations; Functional disorder; Genome; Goals; Hospitalization; Human; Hypertrophy; Immune response; Impairment; In Vitro; Infant; Infection; Iowa; Kidney; Knowledge; Length; Life; Lower Respiratory Tract Infection; Mediating; Molecular; Natural Immunity; Nitric Oxide; Outcome; Parvoviridae; Parvovirus; Parvovirus Infections; Pathogenesis; Pathway interactions; Patients; Physiological; Plasmids; Prevention strategy; Production; Publishing; RNA Viruses; Reactive Oxygen Species; Respiratory Tract Infections; Respiratory syncytial virus; Rhinovirus; Role; Single-Stranded DNA; Surface; System; Testing; Time; Universities; Viral Genome; Viral Proteins; Virion; Virus; Virus Diseases; Wheezing; Work; Xenograft Model; Xenograft procedure; airway epithelium; cell type; clinically relevant; design; effective therapy; experience; gene therapy; in vitro Model; insight; member; positional cloning; public health relevance; research study; respiratory; respiratory infection virus; respiratory virus; response; transmission process; treatment strategy

DESCRIPTION (provided by applicant): Human bocavirus 1 (HBoV1) (genus, Bocavirus; family, Parvoviridae) is a respiratory virus that causes acute respiratory tract infections with wheezing in young children, particularly in infants under two years old. It is commonly associated with other respiratory viruses (with a detection rate of 12-16%) and is the fourth most common respiratory virus after respiratory syncytial virus, adenovirus and rhinovirus in infants under two years old who are hospitalized for the treatment of acute wheezing. Acute HBoV1 infection causes respiratory illness, which can be life-threatening in pediatric patients. At present, there are no effective treatment and prevention strategies for HBoV1 infection, especially in infants with a severe wheeze. HBoV1 has characteristics of an autonomous parvovirus, with a single-stranded DNA genome of approximately 5.5-kb. It productively infects polarized primary human airway epithelium (HAE). To date, we have made much progress in our studies of HBoV1 infection. We have established an HBoV1 reverse genetics system in which HBoV1 virions are produced by transfecting the full-length HBoV1 replicative-form genome into human embryonic kidney (HEK) 293 cells. Infection of polarized primary HAE cultures with these HBoV1 virions results in clear cytopathogenic effects, manifested by the disruption of epithelial integrity. We also demonstrated that progeny HBoV1 virions can infect polarized primary HAE cultures at a multiplicity of infection as low as 0.001 viral genome copies/cell when applied to the apical surface. However, the mechanisms underlying the HBoV1-caused loss of epithelial integrity have not been studied. In addition, the synergistic effects of HBoV1 when co-infected with other respiratory viruses are unknown. Our long-term goal is to use polarized HAE cultures and human bronchial xenografts (physiologically relevant to HBoV1 infection) to identify the key molecular mechanisms underlying HBoV1 replication and the resulting destruction of epithelial integrity. Over the past few years, we have accumulated a great deal of knowledge regarding how host cells respond to parvovirus infection and how the virus induces cell death. For the study proposed herein, we will collaborate with the Center for Gene Therapy at the University of Iowa and use polarized primary HAE cultures and human bronchial xenografts to: i) elucidate the mechanisms by which HBoV1 destroys airway epithelial integrity, reveal the associated morphological and ultrastructural changes of the infected airway epithelium, and identify which cell types of airway epithelium are infected by HBoV1 and which viral proteins mediate the epithelial damage during HBoV1 infection; and ii) determine the synergetic effects of HBoV1 in both polarized HAE cultures and human bronchial xenografts co-infected with other respiratory viruses.

描述（由申请人提供）：人类博卡病毒 1 型 (HBoV1)（博卡病毒属；细小病毒科）是一种呼吸道病毒，可引起幼儿（特别是两岁以下婴儿）的急性呼吸道感染并伴有喘息。它通常与其他呼吸道病毒相关（检出率为12-16%），是因急性喘息住院治疗的两岁以下婴儿中继呼吸道合胞病毒、腺病毒和鼻病毒之后第四常见的呼吸道病毒。急性 HBoV1 感染会导致呼吸道疾病，可能危及儿科患者的生命。目前，针对HBoV1感染，尤其是严重喘息的婴儿，尚无有效的治疗和预防策略。 HBoV1 具有自主细小病毒的特征，其单链 DNA 基因组约为 5.5 kb。它能有效地感染极化的初级人类气道上皮 (HAE)。迄今为止，我们在 HBoV1 感染的研究方面取得了很大进展。我们建立了 HBoV1 反向遗传学系统，其中通过将全长 HBoV1 复制型基因组转染至人胚肾 (HEK) 293 细胞中来产生 HBoV1 病毒颗粒。用这些 HBoV1 病毒粒子感染极化的原代 HAE 培养物会导致明显的细胞病变效应，表现为上皮完整性的破坏。我们还证明，当应用于顶端表面时，子代 HBoV1 病毒体可以以低至 0.001 个病毒基因组拷贝/细胞的感染复数感染极化的原代 HAE 培养物。然而，HBoV1 导致上皮完整性丧失的机制尚未得到研究。此外，HBoV1与其他呼吸道病毒共同感染时的协同作用尚不清楚。我们的长期目标是使用极化 HAE 培养物和人支气管异种移植物（与 HBoV1 感染生理相关）来确定 HBoV1 复制和由此导致的上皮完整性破坏的关键分子机制。在过去的几年里，我们积累了大量关于宿主细胞如何响应细小病毒感染以及病毒如何诱导细胞死亡的知识。对于本文提出的研究，我们将与爱荷华大学基因治疗中心合作，使用极化原代 HAE 培养物和人支气管异种移植物来：i) 阐明 HBoV1 破坏气道上皮完整性的机制，揭示相关的形态学和受感染气道上皮的超微结构变化，并确定哪些气道上皮细胞类型被 HBoV1 感染以及哪些病毒蛋白在感染过程中介导上皮损伤HBoV1 感染； ii) 确定 HBoV1 在极化 HAE 培养物和与其他呼吸道病毒共感染的人支气管异种移植物中的协同效应。