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

描述(申请人提供)：小鼠mshi突变(男性不育和组织不相容)定义了一个独特的次要组织相容(H)基因座的例子，它可能是由一个高度保守的基因突变引起的，似乎介导了一种不依赖于细胞毒性T淋巴细胞(CTL)的不寻常的同种异体移植排斥机制。我们认为，这些非典型属性使MSHI成为一个特别有价值的模型，它可能显著增加我们目前对微小H基因座如何构成的理解，并可以调节MHC匹配个体的免疫反应。我们最近在微管相关蛋白7基因中发现了mshi特异性缺失，这似乎是突变表型中男性不育的完全原因。在这里，我们建议使用不同的皮肤移植物交换试验来明确地确定突变的Mapap7等位基因(Map7mshi缺失和Map7k.o，一种工程功能丧失的“敲除”突变)是否会在小鼠中产生隐性的、单基因的、抗原丢失的障碍，阻碍移植物的相容。同时开发了一种携带Map 7k.o的同源品系。等位基因，我们发现了与Map7k.o相关的组织相容性的显性抗原获得障碍。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屏障是否是由Map7k.o产生的。突变本身，或者相反，是敲除插入两侧的一个(或多个)基因的结果。这些短期研究有望正式将MSHI定义为单基因移植障碍，并为Matt7k.o提供新的同源小鼠资源。这将有助于未来对这一新的、目前未得到充分利用的H模型进行整体动物、体外细胞和生化分析。重要的是，这些研究将加强CCSU的本科生和硕士水平的研究活动，为学生提供学习免疫遗传学方法、收集和分析数据并正式介绍他们的结果的机会，同时它们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.