Host Genetic Factors to Combat Lassa Hemorrhagic Fever

对抗拉沙出血热的宿主遗传因素

基本信息

批准号：
8573827
负责人：
MICHAEL B OLDSTONE
金额：
$ 44.53万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8573827
关键词：
Affect Africa African Air Alleles Alternative Splicing Animal Model Applications Grants Area Arenavirus Binding Biological Assay Biological Warfare Categories Cell Line Cells Cellular biology Centers for Disease Control and Prevention (U.S.)Cessation of life Code Complex Defect Dendritic Cells Disease Enzymes Europe Evolution Fibroblasts Fruit Functional RNA Generations Genes Genetic Genetic Markers Genetic Polymorphism Genetic Transcription Genetic Variation Grant Hand Heart Human ITGAX gene Immigrant Immune response Immune system Implant In Vitro Individual Infection Laboratories Lassa fever virus Learning Letters Life Link Lymphocytic choriomeningitis virus Measures Mediating Messenger RNA Molecular Molecular Biology Molecular Conformation Mus Mutant Strains Mice Mutate Mutation Natural Selections Nigeria Persons Play Population Post-Translational Protein Processing Predisposition Public Health RNA Interference RNA Splicing Receptor Cell Recombinants Recovery Reporting Resistance Role Sampling Serum Shapes Sierra Leone Skeletal Muscle Source Surrogate Markers Surveys Survivors Technology Testing Tissues Transgenes Transgenic Mice United States Viral Viral Hemorrhagic Fevers Viremia Virus Virus Diseases Virus Receptors Virus Replication Western Africa alpha Dystroglycan biosafety level 4 facility combat design genetic selection genome sequencing glycosylation glycosyltransferase in vitro testing in vivo killings mRNA Expression microbial monocyte mouse model mutant novel prevent promoter public health relevance receptor recombinant virus sugar virus pathogenesis weapons

Affect Africa African Air Alleles Alternative Splicing Animal Model Applications Grants Area Arenavirus Binding Biological Assay Biological Warfare Categories Cell Line Cells Cellular biology Centers for Disease Control and Prevention (U.S.)Cessation of life Code Complex Defect Dendritic Cells Disease Enzymes Europe Evolution Fibroblasts Fruit Functional RNA Generations Genes Genetic Genetic Markers Genetic Polymorphism Genetic Transcription Genetic Variation Grant Hand Heart Human ITGAX gene Immigrant Immune response Immune system Implant In Vitro Individual Infection Laboratories Lassa fever virus Learning Letters Life Link Lymphocytic choriomeningitis virus Measures Mediating Messenger RNA Molecular Molecular Biology Molecular Conformation Mus Mutant Strains Mice Mutate Mutation Natural Selections Nigeria Persons Play Population Post-Translational Protein Processing Predisposition Public Health RNA Interference RNA Splicing Receptor Cell Recombinants Recovery Reporting Resistance Role Sampling Serum Shapes Sierra Leone Skeletal Muscle Source Surrogate Markers Surveys Survivors Technology Testing Tissues Transgenes Transgenic Mice United States Viral Viral Hemorrhagic Fevers Viremia Virus Virus Diseases Virus Receptors Virus Replication Western Africa alpha Dystroglycan biosafety level 4 facility combat design genetic selection genome sequencing glycosylation glycosyltransferase in vitro testing in vivo killings mRNA Expression microbial monocyte mouse model mutant novel prevent promoter public health relevance receptor recombinant virus sugar virus pathogenesis weapons

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项目摘要

DESCRIPTION (provided by applicant): This grant focuses on arenavirus pathogenesis (how viruses cause disease) and why during infection some hosts become severely ill and die while others remain relatively unscathed. Lessons learned from lymphocytic choriomeningitis virus (LCMV) are applied to Lassa fever virus (LASV). LASV infects over 300,000 individuals per year resulting in over 20,000 deaths. LASV has been transported by travelers from Western Africa to the USA. Its potential as a weapon of biowarfare is a source of national concern. LASV is placed on the Category A list of microbial agents. My laboratory identified alpha-dystroglycan (¿-DG) as the host cell receptor for LCMV and LASV and demonstrated preferential localization of ¿-DG on dendritic cells (DCs). We then showed that post- translational modification of ¿-DG by the glycosyltransferase LARGE is absolutely critical for the functional maturation of ¿-DG as a receptor for LASV and LCMV binding, entry and replication. P. Sabeti and colleagues found that mutations in LARGE were common in West African populations of Nigeria and Sierra Leone where LASV infection is endemic, but absent in those populations in West Africa where LASV is not endemic. Polymorphisms of the LARGE allele are present in about 35% of the population where LASV is endemic with about 45% of individuals heterozygous and 16% homozygous for it. Mutations in LARGE are found in LASV survivors. Since high viremia (> 9 logs of plaque forming units [PFU/ml of sera]) is routinely associated with signs of severe LASV infection and death, whereas lower viremia (< 8 logs of PFU/ml or less) with recovery, and the ¿-DG/LARGE complex controls LASV entry and subsequent replication, it is likely that mutations in LARGE shape evolution by favoring survival of the host against this virus. Limiting the amount of virus made provides the host's immune system a window of opportunity to mount an effective anti-LASV immune response thereby preventing severe disease and promoting virus clearance. This grant's purpose is to determine if LARGE mutations are responsible for survival to LASV infection. We will assay cells permissive to LASV infection (DCs/monocytes) that contain LARGE mutations in a functional LARGE enzyme assay, and for virus binding, entry and replication using a LASV Gp/LCMV recombinant virus. The recombinant virus can be used in BSL/2 laboratory and has been shown to bind, enter and replicate in permissive human and mouse cells. Thereafter those DCs and monocytes from individuals having LARGE mutations and displaying altered LASV binding, entry or replication or enzymatic activity will be tested at CDC with wtLASV. The fruits of these studies provide the opportunity to identify forces shaping human evolution in that individuals possessing LARGE mutant alleles are likely selected for resistance to LASV. Further, these studies may also identify surrogate markers to classify susceptible or resistant individuals to this viral infection.

描述（由适用提供）：该赠款重点介绍了Arenaviru的发病机理（病毒引起疾病）以及为什么在感染期间，有些宿主会严重生病，死亡，而其他宿主仍然相对毫发无损。从淋巴细胞绒毛膜炎病毒（LCMV）中学到的经验教训用于LASSA热病毒（LASV）。 LASV感染每年超过300,000个人，导致20,000多人死亡。 LASV已被西非的旅行者运送到美国。它作为生物瓦尔法的武器的潜力是国家关注的根源。 LASV放置在微生物剂列表上。我的实验室确定α-毛线糖（Alpha-dystroglycan）（-dg）为LCMV和LASV的宿主细胞受体，并在树突状细胞（DCS）上表现出优先定位。-DG。然后，我们表明，糖基转移酶对„ -dg的转化后修饰对于oodv和lcmv结合，进入和复制的接收器的功能成熟至少至关重要。萨贝蒂（P. Sabeti）及其同事发现，尼日利亚的西非人口和塞拉利昂（Sierra Leone）在LASV感染是内在的，但在LASV不具有内在的西非人群中不存在。大约35％的LASV是内粒，大约45％的个体杂合子和16％的纯合子，大约35％的人口中存在多态性。在LASV表面上发现了大突变。 Since high viremia (> 9 logs of plaque forming units [PFU/ml of sera]) is routinely associated with signs of severe LASV infection and death, whereas lower viremia (< 8 logs of PFU/ml or less) with recovery, and the ¿ -DG/LARGE complex controls LASV entry and subsequent replication, it is likely that mutations in LARGE shape evolution by favoring survival of the host against this 病毒。限制生成的病毒量为宿主的免疫系统提供了有效的抗LASV免疫反应的机会之窗，从而预防严重疾病并促进病毒清除率。该赠款的目的是确定大突变是否负责LASV感染的生存。我们将测定允许在功能性大酶测定中包含大突变的LASV感染（DC/单核细胞）的细胞，以及使用LASV GP/LCMV重组病毒的病毒结合，进入和复制。重组病毒可用于BSL/2实验室，并已显示在允许的人和小鼠细胞中结合，进入和复制。此后，这些DC和单核细胞来自具有较大突变的个体，并显示LASV结合，进入或复制或酶促活性的改变，将通过WTLASV在CDC上进行测试。这些研究的果实为识别塑造人类进化的力提供了机会，因为拥有大型突变等位基因的个体可能被选择以抵抗LASV。此外，这些研究还可以鉴定替代标记物，以对这种病毒感染进行敏感或抗性个体分类。