Molecular study of mouse viral resistance mechanisms

小鼠病毒抵抗机制的分子研究

• 批准号: 7368033
• 负责人: Michael G Brown
• 金额: $ 38.16万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7368033
• 关键词: Acute; Alleles; Antiviral Agents; Backcrossings; Biochemical; Biological Assay; Breeding; C57BL/6 Mouse; Candidate Disease Gene; Cell-Mediated Cytolysis; Cells; Chromosomes; Class; Complex; Congenic Strain; Consensus; Cytolysis; Cytomegalovirus; Cytomegalovirus Infections; DNA Viruses; Data; Defense Mechanisms; Funding; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Genome; Head; Herpesviridae; Histocompatibility Antigens Class II; Homologous Gene; Host Defense; Human; Hybrids; Immune; Immunity; Inbred BALB C Mice; Individual; Infection; International; Laboratories; Ligands; Link; Location; Mediating; Molecular; Murid herpesvirus 1; Mus; NZW Mouse; Natural Immunity; Natural Killer Cells; Operating System; Personal Satisfaction; Play; Predisposition; Proteins; Publishing; Quantitative Genetics; RNA; Range; Receptor Activation; Recurrence; Research Personnel; Research Proposals; Resistance; Resolution; Role; Staining method; Stains; System; Testing; Time; Uncertainty; Universities; Variant; Viral; Virginia; Virus; Virus Diseases; base; genetic strain; member; pathogen; programs; prospective; receptor; resistance mechanism; trait

NK cells are well known for the capacity to recognize and lyse virus infected cells. Indeed, host defenses in humans and mice that lack NK cell immunity are quite vulnerable to overwhelming and recurrent Herpes virus infections. In C57BL/6 mice, host anti-murine cytomegalovirus (MCMV) immunity requires NK cell expressed Ly49H activation receptor recognition of its virus encoded m157 ligand on MCMV infected cells. We recently showed that NK cells in NZW mice however, control experimental MCMV infection without a role for Ly49H+ NK cells. Herein we have extended our finding in MA/My mice. The MA/My strain is noteworthy since it also displays very effective NK cell-mediated control of MCMV infection, but NK cells in this strain do not express Ly49H receptors. Interestingly, MCMV resistance in MA/My is strongly associated with MHC and also non MHC genes. Thus, we have found that NK cells utilize multiple antiviral control mechanisms that are distinguished by genetic polymorphism. The long-term objective of this research proposal therefore is to understand how genetic variation in host genes can contribute differently in innate immunity, its capacity to rapidly recognize and destroy viral pathogens at early times after infection and the molecular and cellular mechanisms controlling such host defenses. The Specific Aims herein will focus initially on the identification and characterization of host genes that contribute substantially to innate anti-MCMV immunity through classical Mendelian genetics studies. The approach is based on rapid phenotypic characterization of hybrid offspring after experimental MCMV infection and subsequent genome-wide genotypic identification of each individual. Using this high-throughput genetics strategy, chromosome locations will be identified in quantitative genetics strategies, confirmed in interval-specific congenic strains, and subsequently candidate genes will be assessed in prospective molecular and biochemical analyses. To facilitate identification of host virus resistance genes, we will also investigate NK cell recognition of virus infection in cellular cytotoxicity assays and virus strain variant selection will also be used to understand innate host defenses mechanistically. While NK cells employ multiple defense mechanisms to control viral pathogens immediately after infection and before adaptive immunity is competent, our studies will no doubt have important implications for human innate defenses in CMV and potentially other virus infections.

众所周知，NK 细胞具有识别和裂解病毒感染细胞的能力。事实上，主机防御 缺乏 NK 细胞免疫力的人类和小鼠很容易感染压倒性和复发性疱疹 病毒感染。在 C57BL/6 小鼠中，宿主抗鼠巨细胞病毒 (MCMV) 免疫需要 NK 细胞 在 MCMV 感染细胞上表达 Ly49H 激活受体识别其病毒编码的 m157 配体。 然而，我们最近发现 NZW 小鼠中的 NK 细胞可以控制实验性 MCMV 感染，但没有发挥作用 对于 Ly49H+ NK 细胞。在此，我们在 MA/My 小鼠中扩展了我们的发现。 MA/My 菌株值得注意 因为它还表现出非常有效的 NK 细胞介导的 MCMV 感染控制，但该菌株中的 NK 细胞不 不表达 Ly49H 受体。有趣的是，MA/My 中的 MCMV 耐药性与 MHC 密切相关 以及非 MHC 基因。因此，我们发现 NK 细胞利用多种抗病毒控制机制 是通过遗传多态性来区分的。因此，本研究计划的长期目标 是了解宿主基因的遗传变异如何对先天免疫做出不同的贡献，其能力 在感染后早期快速识别和消灭病毒病原体以及分子和细胞 控制此类宿主防御的机制。本文的具体目标首先集中于识别 以及对先天性抗 MCMV 免疫有重大贡献的宿主基因的表征 经典孟德尔遗传学研究。该方法基于杂种的快速表型表征 实验性 MCMV 感染和随后的全基因组基因型鉴定后的后代 个人。使用这种高通量遗传学策略，染色体位置将在 定量遗传学策略，在特定区间的同源菌株中得到证实，随后候选 基因将在前瞻性分子和生化分析中进行评估。方便识别主机 病毒抗性基因，我们还将研究NK细胞识别病毒感染的细胞毒性 检测和病毒株变体选择也将用于了解宿主的先天防御 机械地。 NK细胞利用多种防御机制立即控制病毒病原体 在感染后和适应性免疫发挥作用之前，我们的研究无疑将具有重要意义 对巨细胞病毒和其他潜在病毒感染的人类先天防御的影响。