RESPONSES OF INBRED MICE TO ECTROMELIA VIRUS

近交系小鼠对兔斑病毒的反应

• 批准号: 2281596
• 负责人: DAVID G BROWNSTEIN
• 金额: $ 22.32万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-01-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2281596
• 关键词: Poxviridae; animal population genetics; complement; disease /disorder proneness /risk; epizootiology; genetic mapping; genetic strain; host organism interaction; inbreeding; laboratory mouse; natural killer cells; tissue mosaicism; veterinary medicine; virus cytopathogenic effect; virus replication

DESCRIPTION (Adapted from Applicant's Abstract): Mousepox is an important viral disease of laboratory mice with remarkably varied symptomatology based on the presence or absence of multiple resistance genes. The long term objectives of this research are to identify, localize and clone the major mousepox resistance genes, to determine their mechanisms of action, and to devise strategies based on transgenic technology to protect susceptible strains from ectromelia virus. For the major mousepox resistance genes, this application proposes to complete development of congenic strains, to continue defining mechanisms of action, to complete chromosomal localization, and to begin fine mapping. Resistance is determined prior to the onset of specific immunity, requires natural killer (NK) cells, and one resistance gene, RMP-1, maps near a complex of genes, NKR-P1, including that which encodes NK 1.1, expressed solely by NK cells. To investigate the role of NK cells in resistance, mixed chimeras will be produced which carry NK cells from resistant NK 1.1+ (Rmp-1r) and susceptible NK 1.1- congenic donors. Three methods will be used to produce mixed chimeras, parabiosis, adoptive transfer of adult spleen cells to infants, and radiation bone marrow chimeras. The method that best demonstrates transfer of resistance to the susceptible congenic partner will be used. The effect of ablation of NK 1.1+ cells in NK 1.1+/NK1.1- chimeras by anti-NK 1.1 antibody on virus replication will determine if Rmp-1 is expressed by NK 1.1+ cells. If Rmp-1 is expressed by NK 1.1+ cells, a modification of this protocol will be used to determine if other resistance genes are expressed by NK cells. The resistant donors will be F1 hybrids of the Rmp-1r congenic strain and congenic strains for the other resistance genes of interest. Because F1 cells will express both dominant resistance genes and NK 1.1, ablation of NK 1.1+ cells will augment virus replication if the other resistance genes are also expressed by NK cells. To determine if resistance genes that are not expressed by NK cells act through NK cells, levels of NK cell expression of NKR-P1 genes and immune interferon (IFN-gamma), which exhibits sex-biased expression like some resistance genes, will be tested in ectromelia infected congenic strains.The complement system and T cell precursors have also been implicated in resistance to mousepox.To determine if those congenic resistant strains with genes that are not expressed by or through NK cells mediate resistance through complement of T cell precursors. The effects of complement and T cell depletion on virus replication will be examined. The chromosomal location of Rmp-4, the only unlocalized resistance gene, will be determined by multilocus analysis of the Rmp-4 congenic strain and the susceptible congenic partner using probes for mapped non ecotropic proviruses. The four resistance genes will be fine mapped by determining loci on the donor segments of congenic resistant strains and using these loci in linkage analysis with resistance genes in backcross mice.

描述（改编自申请人的摘要）：Mousepox是一个 实验室小鼠的重要病毒疾病，有差异很大 症状学基于多个的存在或不存在 抗性基因。这项研究的长期目标是 识别，定位和克隆主要的摩氏抗性基因， 确定其行动机制，并设计基于策略 关于保护易感性菌株免受的转基因技术 胚胎病毒。 对于主要的摩氏抗性基因，这是 申请建议完整开发先天性菌株，以 继续定义作用机理，以完成染色体 本地化，开始精细的映射。 确定电阻 在特定免疫发作之前，需要自然杀手（NK） 细胞和一个抗性基因RMP-1，在基因复合物附近地图， NKR-P1，包括编码NK 1.1的NK-P1，仅由NK表示 细胞。 为了研究NK细胞在抗性中的作用，混合 将产生嵌合体，该嵌合体携带NK 1.1+的NK细胞 （RMP-1R）和敏感的NK 1.1-先天捐助者。 将三种方法 用于生产混合嵌合体，抛物线病，收养 成人脾细胞向婴儿和辐射骨髓嵌合体。 最能证明电阻转移到该方法的方法 将使用敏感的同类伙伴。 消融的影响 NK 1.1+/nk1.1-在抗NK 1.1嵌合在抗NK 1.1抗体上的NK 1.1+细胞。 病毒复制将确定RMP-1是否由NK 1.1+表示 细胞。 如果RMP-1由NK 1.1+单元表达 该方案将用于确定其他抗性基因是否是 由NK细胞表达。 抗性供体将是F1的杂种 另一种RMP-1R的先天性菌株和先天性菌株 感兴趣的抗性基因。 因为F1单元将同时表达 显性抗性基因和NK 1.1，NK 1.1+细胞的消融将 增强病毒复制如果其他抗性基因也是 由NK细胞表达。 确定抗性基因是否不是 由NK细胞表达通过NK细胞的作用，NK细胞的水平 NKR-P1基因和免疫干扰素的表达（IFN-GAMMA）， 表现出性别偏见的表达，例如某些抗性基因，将是 在胚胎感染的先天性菌株中测试。补体系统 和T细胞前体也与对 摩托福克。确定那些具有基因的抗性菌株是否存在 不受NK细胞表达的介导电阻的表达 通过补充T细胞前体。补充的影响 将检查病毒复制的T细胞耗竭。 这 RMP-4的染色体位置，RMP-4是唯一的非钙化抗性基因， 将通过对RMP-4先生的多焦点分析来确定 使用探针进行映射的菌株和易感的同类伴侣 非生态病毒。 四个电阻基因将很好 通过确定基因座的位点，在抗抗药性的供体部分上 菌株并使用这些基因座在链接分析中具有阻力 反向交叉小鼠中的基因。