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

描述（由申请人提供）：小鼠mshi突变（雄性不育和组织不相容性）定义了一个次要组织相容性(H)位点的独特例子，它可能是由单个高度保守的基因突变引起的，并且似乎介导了一种不寻常的异体移植物排斥机制，该机制不依赖于细胞毒性T淋巴细胞（CTL）。我们相信这些非典型属性使mshi成为一个特别有价值的模型，可以显著地增加我们目前对mhc匹配个体中次要H位点如何结构和调节免疫反应的理解。我们最近在Mtap7基因（微管相关蛋白7）中发现了mashi特异性缺失，这似乎是突变表型中男性不育方面的完全原因。在这里，我们建议使用不同的皮肤移植物交换试验来明确确定突变的Mtap7等位基因（Mtap7mshi缺失和Mtap7k.o）是否会导致基因突变。（一种工程缺失功能的“敲除”突变）可以在小鼠体内产生隐性的、单基因的、抗原缺失的移植物相容性障碍。在开发携带Mtap7k.o基因系的过程中。我们发现了与Mtap7k.o相关的显性抗原获得性组织相容性屏障。突变。因此，我们建议使用另一种皮肤移植交换试验来确定这种显性h -屏障是否由Mtap7k.o产生。突变本身，或者是一个基因（或多个基因）在敲除插入旁边的结果。这些短期研究预计将正式定义mshi为单基因移植屏障，并为Mtap7k.o提供新的同源小鼠资源。这将有助于未来对这种新颖且目前未充分利用的H模型进行全动物、体外细胞和生化分析。重要的是，这些研究将通过为学生提供学习免疫遗传学方法、收集和分析数据以及对结果进行正式展示的机会，从而加强CCSU本科生和硕士水平的研究活动，同时有助于将独特的组织相容性模型带入功能和分子分析的主流。

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.