Do genetic variants of Mtap7 create single-gene barriers to tissue-graft compatib

Mtap7 的遗传变异是否会造成组织移植相容性的单基因障碍

• 批准号: 8313899
• 负责人: THOMAS R KING
• 金额: $ 7.16万
• 依托单位: CENTRAL CONNECTICUT STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-15 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8313899
• 关键词: Alleles; Alloantigen; Allogenic; Animals; Antigens; Binding; Biochemical; Biological Assay; CD4 Positive T Lymphocytes; Congenic Mice; Congenic Strain; Connecticut; Cytotoxic T-Lymphocytes; Data Analyses; Defect; Development; Disease; Engineering; Future; Genes; Genetic; Genetic Structures; Helper-Inducer T-Lymphocyte; Histocompatibility; Homozygote; I-antigen; Immune response; Immunogenetics; Immunotherapy; In Vitro; Individual; Investigation; Knock-out; Learning; Life; Link; Mainstreaming; Male Sterility; Mediating; Methods; Microtubule-Associated Proteins; Minor; Modeling; Molecular Analysis; Mouse Strains; Mus; Mutation; Phenotype; Research; Research Activity; Resources; Skin Transplantation; Skin graft; Speed; Structure; Students; Tail; Testing; Tissue Grafts; Tissue Transplantation; Transplantation; Universities; Variant; allograft rejection; bacterial H antigen; congenic; genetic variant; in vivo; loss of function; mutant; novel; positional cloning; response

DESCRIPTION (provided by applicant): The mouse mshi mutation (for male sterility and histoincompatibility) defines a unique example of a minor histocompatibility (H) locus, one that may have resulted from mutation of a single, highly-conserved gene, and that appears to mediate an unusual allograft rejection mechanism that is independent of cytotoxic T lymphocytes (CTL). We believe that these atypical attributes make mshi an especially valuable model that may add significantly to our current understanding of how minor H loci may be structured and can modulate an immune response among MHC-matched individuals. We have recently identified a mshi-specific deletion in the Mtap7 gene (for microtubule- associated protein 7) that appears to be fully responsible for the male-sterility aspect of the mutant phenotype. Here we propose to use distinct skin-graft-exchange assays to explicitly determine whether mutant Mtap7 alleles (the Mtap7mshi deletion and Mtap7k.o., an engineered loss-of-function "knock out" mutation) can generate recessive, single-gene, antigen-loss barriers to graft compatibility in mice. While developing a congenic line that carries the Mtap7k.o. allele, we found a dominant, antigen-gain barrier to histocompatibility associated with the Mtap7k.o. mutation. We therefore propose to use another skin-graft-exchange assay to determine whether this dominant H-barrier is generated by the Mtap7k.o. mutation itself, or is instead the result of a gene (or genes) that flank the knock-out insertion. These short-term investigations are anticipated both to formally define mshi as a single- gene transplantation barrier and to provide new congenic mouse resources for Mtap7k.o. that will facilitate the future whole animal, in vitro cellular, and biochemical analysis of this novel and currently-underutilized H model. Importantly, these investigations will enhance undergraduate and master's-level research activity at CCSU by providing students with opportunities to learn the methods of immunogenetics, to collect and analyze data, and to make formal presentations of their results while they help to bring a unique histocompatibility model into the mainstream of functional and molecular analysis.

描述（由申请人提供）：小鼠mshi突变（雄性不育和组织不相容性）定义了次要组织相容性（H）基因座的一个独特实例，该基因座可能由单个高度保守基因的突变引起，并且似乎介导不依赖于细胞毒性T淋巴细胞（CTL）的不寻常的同种异体移植物排斥机制。我们认为，这些非典型的属性使mshi一个特别有价值的模型，可能会显着增加我们目前的理解，如何小H基因座可能是结构化的，可以调节MHC匹配的个人之间的免疫反应。 我们最近已经鉴定了Mtap 7基因中的mshi特异性缺失（微管相关蛋白7），其似乎完全负责突变体表型的雄性不育方面。在这里，我们提出使用不同的皮肤移植物交换测定来明确地确定突变Mtap 7等位基因（Mtap 7 mshi缺失和Mtap7k.o.，工程化的功能丧失“敲除”突变）可以在小鼠中产生对移植物相容性的隐性、单基因、抗原丧失屏障。在开发携带Mtap7k.o的同类系的同时。等位基因，我们发现了一个显性的，抗原增益障碍与Mtap7k.o的组织相容性。突变因此，我们建议使用另一种皮肤移植物交换试验来确定这种主要的H-屏障是否由Mtap7k.o产生。突变本身，或者替代地是侧翼敲除插入的基因（或多个基因）的结果。 预期这些短期研究将正式将mshi定义为单基因移植屏障，并为Mtap7k.o提供新的同类小鼠资源。这将有助于未来的整体动物，在体外细胞，和生化分析，这种新的和目前未充分利用的H模型。重要的是，这些研究将为学生提供学习免疫遗传学方法、收集和分析数据以及正式展示结果的机会，从而加强CCSU的本科生和硕士研究活动，同时有助于将独特的组织相容性模型纳入功能和分子分析的主流。