THYMUS LEUKEMIA GENES AND ANTIGENS

胸腺白血病(TL)抗原氨基酸序列的研究 相信TL蛋白可能具有一种抗原的假设 结合裂隙类似于在聚集性HLA-A2 I类上发现的结合裂隙 糖蛋白。类似于人类白细胞抗原A2，但以一种受限得多的方式，TL 分子可能能够结合和呈递抗原。在这 背景，最近的报道表明TL是抗原提呈物 T细胞受体(TCR)分子的配体。因此， 这项提议的长期目标是理解 I类基因和蛋白的TL家族的生物学功能 以及调节基因的分子机制 组织特异性表达。 具体地说，转基因小鼠将被生产出来并用于研究 H-2kb启动子驱动TL的表达模式。首字母 实验证明，这种基因在这样的小鼠中表达。 与内源性H-2第I类蛋白相似。抗TCR抗体， 将首先使用CD4和CD8分子对转基因免疫进行染色 细胞，以确定TL的异位表达是否导致 T细胞分化途径的扰动。从免疫学上讲， 将评估TL作为移植的能力 抗原和使用病毒模型的CTL限制元件。正性 结果将允许克隆效应器T细胞和 检查他们的TCR分子。这样的结果可以产生一个 提示TL在体内与之相互作用的配体以及 TL可能的功能作用。这些实验还可以形成 为今后的结构功能研究奠定基础。实验涉及到 推测与TL多肽反应的新型单抗抗体的分析 来自非剪接TL转录本的以及鉴定 参与剪接的TL启动子元件和组织特异性TL 表达将继续。

A study of the thymus-leukemia (TL) antigens' amino acid sequence lends credence to the hypothesis that TL proteins may possess an antigen binding cleft similar to that found on the paradigmatic HLA-A2 class I glycoprotein. Like HLA-A2, but in a far more restricted manner, the TL molecules may be capable of binding and presenting antigen. In this context, recent reports have implicated TL as the antigen presenting ligand for gamma/delta T cell receptor (TcR) molecules. Thus, the long-term objectives of this proposal are to understand both the biological function of the TL family of class I genes and proteins as well as the molecular mechanisms which regulate the genes' tissue-specific expression. Specifically, transgenic mice will be produced and used to study the pattern of expression of TL driven by the H-2Kb promoter. Initial experiments have demonstrated that the gene is expressed in such mice similar to endogenous H-2 class I proteins. Antibodies against TcR, CD4, and CD8 molecules will first be used to stain transgenic immune cells in order to determine whether ectopic expression of TL results in the perturbation of T cell differentiative pathways. Immunologically, TL will be assessed for its ability to serve as a transplantation antigen and a CTL restriction element using virus models. Positive results will allow for the cloning of the effector T cells and an examination of their TcR molecules. Such results can yield an indication as to the ligand with which TL interacts in vivo as well as TL's possible functional role. These experiments can also form the basis for future structure-function studies. Experiments involving the analysis of novel mAb antibodies putatively reacting with TL peptides derived from unspliced TL transcripts as well as the identification of TL promoter elements involved in splicing and tissue-specific TL expression will be continued.