Transcriptomics of immunity and disease in African Fruit Bats- important zoonotic reservoirs

非洲果蝠——重要的人畜共患病宿主的免疫和疾病的转录组学

• 批准号: 9243490
• 负责人: Kenneth A Field
• 金额: $ 22.02万
• 依托单位: BUCKNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243490
• 关键词: Acute; Address; Affect; African; Age; Age Factors; Antiparasitic Agents; Antiviral Agents; Asians; Blood; Cell Culture Techniques; Cells; Characteristics; Chiroptera; Chronic; Comparative Physiology; Coronaviridae; Coronavirus; Cytotoxic T-Lymphocytes; Disease; Disease Management; Ebola virus; Ecology; Environment; Etiology; Family Pteropodidae; Female; Filoviridae; Filovirus; Food; Fruit; Funding; Future; Gene Expression; Gene Expression Profile; Genes; Geography; Goals; Habitats; Health; Heterogeneity; Human; Immune; Immune response; Immunity; Immunocompetence; Immunologic Surveillance; Immunologics; Immunologist; Immunology; Infection; Interferon Type II; Interferon-alpha; Interferons; International; Knowledge; Malaria; Modification; Myelogenous; Oral; Orthomyxoviridae; Outcome; Paramyxoviridae; Paramyxovirus; Parasitemia; Parasites; Pathway interactions; Pattern; Pattern recognition receptor; Population; Process; Program Research Project Grants; Proteins; Regulatory Pathway; Regulatory T-Lymphocyte; Reproductive History; Research; Risk; Role; Salivary Glands; Sampling; Seasons; Sex Factors; Signal Pathway; Site; South Sudan; Spleen; Suppressor-Effector T-Lymphocytes; Surveys; Swab; Taxonomy; Testing; Tissue Sample; Tissue-Specific Gene Expression; Tissues; Toll-like receptors; Training; United States National Institutes of Health; Universities; Variant; Viral; Viral Load result; Viral reservoir; Viremia; Virus; Virus Diseases; Virus Shedding; Weather; Work; Zoonoses; anthropogenesis; antiviral immunity; base; burden of illness; co-infection; comparative; design; differential expression; disease transmission; environmental change; experience; immune function; improved; male; novel; pandemic disease; pathogen; programs; reproductive; response; sex; transcriptome; transcriptome sequencing; transcriptomics

ABSTRACT In line with the funding goals of the NIH and the objectives of the R21 research program, this project uses the power of transcriptomics to understand bat immune competence in relation to viral infection in a natural, variable environment. This project will be jointly led at Bucknell University by co-PD/PIs Dr. DeeAnn Reeder (internationally recognized expert in bat disease and comparative physiology) and Dr. Ken Field (classically trained immunologist with expertise in applying transcriptomic approaches to bat disease ecology). The goals of this work are to explore how intrinsic (age, sex, reproductive condition, current disease status) and extrinsic (seasonal shifts in weather and food availability) factors underlie immunological variation in African fruit bats, reservoirs for viruses of pandemic potential (including Ebola) that are becoming increasingly associated with people due to habitat modification. While important progress has been made in recent years in understanding bat immunity, much of this has been in cell culture or from limited sampling, largely from SE Asian and Australian bats; this study will fill this taxonomic and geographic gap and transform our understanding of variation in antiviral immunity by examining immune processes in the real world. To perform this work, male and female foraging bats will be collected at field sites in South Sudan during both the rainy and dry season. For Specific Aim 1, spleen tissue samples will be used to determine the differential expression (Illumina HiSeq 4000 platform and Trinity analysis pipeline) of genes involved in immune function, with an emphasis on antiviral immunity. Findings will be confirmed in subsequent qPCR studies and will be used to test the recently proposed hypothesis that bat antiviral gene expression is “always on”, which may be related to reservoir capacity. Relationships found between intrinsic and extrinsic factors and immune gene expression will be used to describe periods of low antiviral immunity, which may increase spillover risk. For Specific Aim 2, gene expression in relation to diseased state will be analyzed for bats with exceptionally high malarial parasite (Hepatocystis) loads or with high viral loads (surveying filoviruses, coronaviruses, paramyxoviruses and orthomyxoviruses), compared to matched controls. For genes with differential expression, qPCR will be used to look for similar changes in other tissues, matched to viral findings (e.g., high viral load from oral swabs will prompt gene expression examination in salivary glands). Relationships between gene expression and disease state will be interpreted in the context of the influence of co-infection (malaria) and of viral infection on antiviral mechanisms. If our proposed specific aims are achieved, we will significantly enhance our understanding of bat immunity and the factors that influence it under natural conditions. This will improve our ability to predict when viral spillovers may be more likely and how changing environmental conditions, including the anthropogenic alteration of natural landscapes, may alter disease processes.

摘要 根据NIH的资助目标和R21研究计划的目标，该项目使用 转录组学的力量，以了解蝙蝠的免疫能力与病毒感染的自然， 多变的环境该项目将由巴克内尔大学的共同PD/PI博士DeeAnn里德共同领导 （国际公认的蝙蝠疾病和比较生理学专家）和肯菲尔德博士（经典 训练有素的免疫学家，具有将转录组学方法应用于蝙蝠疾病生态学的专业知识）。的目标 这项工作的目的是探讨内在（年龄，性别，生殖条件，目前的疾病状况）和外在 （天气和食物供应的季节性变化）因素是非洲果蝠免疫变异的基础， 具有大流行潜力的病毒（包括埃博拉病毒）的储存库， 人类由于栖息地的改变。虽然近年来在了解 蝙蝠的免疫力，其中大部分是在细胞培养或从有限的采样，主要是从东南亚和 澳大利亚蝙蝠;这项研究将填补这一分类和地理空白，并改变我们对 通过检查真实的世界中的免疫过程来研究抗病毒免疫的变化。 为了完成这项工作，雄性和雌性觅食蝙蝠将在南苏丹的野外收集， 无论是雨季还是旱季。对于特定目标1，脾组织样本将用于确定 免疫相关基因的差异表达（Illumina HiSeq 4000平台和Trinity分析管道） 功能，重点是抗病毒免疫。结果将在随后的qPCR研究中得到证实， 将被用来测试最近提出的假设，蝙蝠抗病毒基因的表达是“永远”， 可能与水库容量有关。内在和外在因素与免疫功能之间的关系 基因表达将被用来描述抗病毒免疫力低的时期，这可能会增加溢出风险。 对于特定目标2，将分析与疾病状态相关的基因表达，以获得异常的蝙蝠。 高疟疾寄生虫（肝囊肿）载量或高病毒载量（调查丝状病毒，冠状病毒， 副粘病毒和正粘病毒）。对于具有差异的基因， 表达，qPCR将用于寻找其他组织中的类似变化，与病毒发现相匹配（例如，高 来自口腔拭子的病毒载量将提示唾液腺中的基因表达检查）。之间的关系 基因表达和疾病状态将在合并感染（疟疾）影响的背景下进行解释 以及病毒感染对抗病毒机制的影响。如果我们提出的具体目标得以实现， 加强我们对蝙蝠免疫力的了解，以及在自然条件下影响蝙蝠免疫力的因素。这将 提高我们预测何时更有可能发生病毒溢出以及环境如何变化的能力 各种条件，包括人为改变自然景观，可能会改变疾病进程。