ANALYSIS OF H-2 MUTANT STRAINS

这一建议继续了我们对I类分子及其如何突变的研究。 编码这些分子的基因会影响T细胞的识别。一 解释H-2Kbm突变基因多碱基变化的机制 就是基因转化。Q10基因具有突变株H-2Kbm1中的碱基 基因，并被认为是这种突变的供体基因。我们会 产生针对QA抗原的CTL，包括那些不正常表达的抗原 (如Q10)，并确定这些抗原与 H-2Kbm分子(和其他I类分子)。干扰素已经被证明是 上调I类抗原的表达。我们将演示 这种干扰素不会上调某些QA抗原。使用域 重组QA/I类抗原，我们将确定I类基因的区域 这允许干扰素的诱导性，并决定了 干扰素增强CTL活性。QA和TL抗原的结构 与I类分子H-2K、D和L非常相似。 然而，前一种分子并不能作为限制因素。 抗原特异性CTL。由于结构域改组后的QA和TL抗原 在相对较高密度的转基因L细胞上表达，我们将测试 这些分子作为靶分子发挥作用的能力 抗原特异性H-2限制性CTL。在协作研究中，这些QA 和Tla外显子改组基因将被引入小鼠卵子和 转基因小鼠将接受测试，测试它们将这些分子用作 抗原特异性CTL的限制因素。定点突变 MHC基因的研究有助于确定哪些结构影响CTL 承认。我们将确定抗H-2LD CTL的良好特异性 通过测试它们对几个H-2LD突变体的活性来克隆。在……里面 合作研究，T细胞受体基因的结构将是 下定决心。碳水化合物在MHC分子影响CTL中的作用 识别将被确定，删除整个 分子的细胞质部分。最后，它的结构组件 将对VSV进行分析，以确定1)病毒编码的分子 被CTL识别和2)具有不同H-2等位基因的小鼠 对不同的病毒成分做出反应。

This proposal continues our studies on class I molecules and how mutation of the genes that encode these molecules affects T cell recognition. One mechanism to explain the multiple base changes seen in H-2Kbm mutant genes is gene conversion. The Q10 gene has the bases found in the mutant H-2Kbm1 gene and has been proposed to be the donor gene for this mutation. We will generate CTL against Qa antigens, including those not normally expressed (such as Q10), and determine the relationship between these antigens and H-2Kbm molecules (and other class I molecules). Interferon has been shown to up-regulate the expression of class I antigens. We will demonstrate that interferon does not up-regulate some Qa antigens. Using domain shuffled Qa/class I antigens, we will determine the region of class I genes that allows for interferon inducibility and determine the role of interferon in augmenting CTL activity. The structure of Qa and TL antigens is very similar to that of the class I molecules, H-2K, D, and L. Nevertheless, the former molecules do not serve as restricting elements for antigen specific CTL. Since domain shuffled Qa and TL antigens are expressed on transfected L cells at relatively high density, we will test the ability of these molecules to function as target molecules for antigen-specific H-2 restricted CTL. In collaborative studies, these Qa and Tla exon shuffled genes will be introduced into mouse ova and the transgenic mice will be tested for their ability to use these molecules as restricting elements for antigen-specific CTL. Site specific mutagenesis of MHC genes allows for a determination of what structures affect CTL recognition. We will determine the fine specificity of anti-H-2Ld CTL clones by testing their activity against several H-2Ld mutants. In collaborative studies, the structure of the T-cell receptor genes will be determined. The role of carbohydrate on MHC molecules in affecting CTL recognition will be determined, as will the affect of removing the entire cytoplasmic portion of the molecule. Finally, the structural components of VSV will be analyzed to determine 1) which molecules encoded by the virus are recognized by CTL and 2) whether mice with differing H-2 alleles respond to different viral components.