A model for developmental IFN gene regulation in the virus-infected fetus

病毒感染胎儿发育干扰素基因调控模型

• 批准号: 7981268
• 负责人: RAYMOND ROWLAND
• 金额: $ 43.32万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7981268
• 关键词: Active Learning; Acute; Animal Model; Antibodies; Antibody Formation; Antiviral Agents; Antiviral Response; Arterivirus; Biological; Biological Models; Data; Development; Disease; Educational process of instructing; Embryo; Environmental Risk Factor; Equipment and supply inventories; Etiology; Family; Family suidae; Fetus; Gene Expression; Gene Expression Regulation; Genes; Health; Human; Immune response; Immunity; Immunocompetent; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Interferon Receptor; Interferon Type I; Interferon Type II; Interferons; Laboratories; Link; Maintenance; Measures; Medicine; Modeling; Molecular; Molecular Profiling; Natural Immunity; Neonatal; Normal tissue morphology; Outcome; Pathogenesis; Pathway interactions; Pattern; Pharmaceutical Preparations; Placenta; Porcine Reproductive and Respiratory Syndrome; Porcine respiratory and reproductive syndrome virus; Pregnancy; Regulation; Research; Reverse Transcriptase Polymerase Chain Reaction; Students; Study models; T-Lymphocyte; Techniques; Testing; Thymus Gland; Time; Tissues; Viremia; Virus; Virus Diseases; Virus Replication; comparative; cytokine; fetal; fetal infection; functional group; human disease; implantation; innovation; pig genome; pregnant; public health relevance; receptor; receptor expression; response; skills

DESCRIPTION (provided by applicant): The interferons (IFNs) are a multi-functional group of cytokines that participate in the implantation of the early embryo, maintenance of a successful pregnancy, initiation of innate immunity and regulation of adaptive immune responses. A recent gene inventory of the pig genome identified 39 type I, 1 type II and 2 type III IFN genes. The biological significance of a large number of genes from the same family suggests that IFN expression is developmentally regulated, including the up-regulated expression of specific classes, subclasses and subtypes during viral infection of the fetus. The purpose of the proposed project is to profile the expression of IFN genes in their receptors in healthy and virus-infected fetuses. The model system incorporates the natural infection of the late-gestation pig fetus with an arterivirus, porcine reproductive and respiratory syndrome virus (PRRSV). Aim 1 is to characterize the expression of IFN classes, subclasses and subtypes in the immunocompetent pig fetus. The hypothesis being tested is that specific IFN genes are preferentially expressed in the late gestation fetus and the pattern of gene expression is tissue specific. Under this aim, a real-time RT-PCR array is used to measure the expression of the individual porcine IFNs and their receptors. Expression will be measured in fetal, interface (placenta) and maternal compartments at 75, 85, 92, 100 and 110 days of the 114 day gestation period. Aim 2 is to measure alterations in fetal IFN gene expression during virus infection. Natural infection of the pregnant porcine dam with PRRSV at 85 to 90 days gestation leads to fetal infection. Two isolates that possess two different endpoints are used for infection. IFN and IFN receptor expression will be measured at 112 days of gestation. The pattern of gene expression will be correlated with viremia and PRRSV-specific antibody. The results will identify candidate IFNs that are involved in protecting the fetus or may be involved in fetal rejection. Aim 3 is the participation of pre-veterinary (prevet) undergraduate students into research. The experiential learning activities will teach undergraduate students research skills related to the development and use of a comparative animal model and basic molecular laboratory techniques. PRRSV infection of the late-gestation fetus provides a relevant model system for dissecting the interaction between a virus and its natural host. Another important innovation is the analysis of IFN gene expression in the immunocompetent fetus, which will provide an important comparative model for studies of human fetal infections, their consequences and potential therapies. PUBLIC HEALTH RELEVANCE: This project is the first-of-its-kind analysis of IFN and IFN-gene expression in the immunocompetent pig fetus. The model provides the means to test therapies and other treatments on fetal immunity and identify expression patterns involved in defense and pathogenesis.

描述（申请人提供）：干扰素（IFN）是一组多功能细胞因子，参与早期胚胎的植入、成功妊娠的维持、先天免疫的启动和适应性免疫反应的调节。最近的猪基因组基因库鉴定出 39 个 I 型、1 个 II 型和 2 个 III 型 IFN 基因。来自同一家族的大量基因的生物学意义表明，IFN表达受到发育调节，包括胎儿病毒感染期间特定类、亚类和亚型的表达上调。该项目的目的是分析健康胎儿和病毒感染胎儿的受体中干扰素基因的表达。该模型系统结合了妊娠晚期猪胎儿对动脉病毒、猪繁殖与呼吸综合征病毒（PRRSV）的自然感染。目标 1 是表征免疫活性猪胎儿中 IFN 类别、亚类和亚型的表达。正在测试的假设是特定的干扰素基因优先在妊娠晚期胎儿中表达，并且基因表达模式是组织特异性的。在此目标下，使用实时 RT-PCR 阵列来测量个体猪 IFN 及其受体的表达。将在 114 天妊娠期的第 75 天、85 天、92 天、100 天和 110 天测量胎儿、界面（胎盘）和母体隔室中的表达。目标 2 是测量病毒感染期间胎儿 IFN 基因表达的变化。怀孕母猪在妊娠 85 至 90 天时自然感染 PRRSV 会导致胎儿感染。具有两个不同终点的两种分离株用于感染。干扰素和干扰素受体的表达将在妊娠 112 天时进行测量。基因表达模式将与病毒血症和PRRSV特异性抗体相关。结果将鉴定出参与保护胎儿或可能参与胎儿排斥反应的候选干扰素。目标 3 是兽医预科（prevet）本科生参与研究。体验式学习活动将教授本科生与比较动物模型的开发和使用以及基本分子实验室技术相关的研究技能。妊娠晚期胎儿的 PRRSV 感染为剖析病毒与其自然宿主之间的相互作用提供了一个相关的模型系统。另一项重要的创新是对免疫健全胎儿中干扰素基因表达的分析，这将为研究人类胎儿感染、其后果和潜在疗法提供重要的比较模型。 公共健康相关性：该项目是首次对具有免疫能力的猪胎儿中的 IFN 和 IFN 基因表达进行分析。该模型提供了测试胎儿免疫疗法和其他治疗方法并识别参与防御和发病机制的表达模式的方法。