IMMUNOGLOBULIN-RELATED T LYMPHOCYTE SURFACE RECEPTORS

这项研究旨在获得详细的信息， 定义的抗原特异性表面组分的结构性质 鼠和灵长类T细胞系。 总体目标是获得详细的 关于生物化学和分子生物学特性的信息 T细胞表面成分，这些成分在血清学上与 免疫球蛋白-VH 区域标记（V不），这将允许直接 与其他可能的T细胞受体（例如表面复合物）的比较 受主要组织相容性复合体的限制。 具体目标是： (1)来描述V不- 结构上相关的T细胞分子。 的 这里的主要方法是开始用大量的同质的， 体外生长的T细胞系（大于50克），并制备 免疫亲和层析法。 具体条件 将使用肽键的切割， 通过高效液相色谱法。 (2)受限V不- 相关 由T细胞系70-N2表达和合成的决定簇被遗传 在人类中是常染色体显性遗传。 康贝特人将以 指定该标记的基因的遗传连锁。 (3)为了确保 获得完整的序列信息，我们将接近 V问题不- 携带我们定义的细胞系的T细胞受体， 分子生物学技术。 我们将用我们的特异性抗血清 从我们分离的带有新生多肽链的完整多聚核糖体 细胞系，分离mRNA并产生cDNA，然后克隆， cDNA的测序。 另一种策略是建造 使用总mRNA和表达载体pUC 8的cDNA文库， 允许合成抗原产物。 我们将筛选cDNA文库 用缺口翻译的DNA探针直接针对假定的T细胞表面 受体。 然后用我们的抗血清筛选阳性克隆， 确定我们产生的抗V不- 相关产品 最近与假定的T细胞受体基因的产物反应 测序 （简体中文）

This research is directed towards obtaining detailed information on the structural properties of the antigen-specific surface component of defined murine and primate T-cell lines. The overall goal is to obtain detailed information regarding the biochemical and molecular biological properties of T-cell surface components which are serologically related to immunoglobulin-VH region markers (VT), which will allow direct comparison with other possible T-cell receptors such as surface complexes restricted by the major histocompatibility complex. The specific aims are: (1) to characterize the VT-related T-cell molecules structurally. The major approach here will be to begin with large quantities of homogenous, in vitro-grown T-cell lines (greater than 50 grams) and to prepare the molecules by immune affinity chromatography. Conditions for specific cleavage of peptide bonds will be used and the fragments will be separated by high performance liquid chromatography. (2) The restricted VT-related determinant expressed and synthesized by the T-cell line 70-N2 is inherited as an autosomal dominant within human populations. We will determine the genetic linkage of the gene specifying this marker. (3) In order to ensure that complete sequence information is obtained, we will approach the problem of the VT-bearing t-cell receptor of our defined lines using the techniques of molecular biology. We will use our specific antisera to isolate intact polyribosomes bearing nascent polypeptide chains from our cell lines, isolate mRNA and produce cDNA followed by cloning and sequencing of the cDNA. An alternative strategy will be the construction of a cDNA library using total mRNA and the expression vector pUC8 which allows synthesis of antigenic products. We will screen the cDNA library with nicktranslated DNA probes directed against putative T-cell surface receptors. The positive clones will then be screened with our antisera to determine whether the antisera we have generated to VT-related products react with the products of the putative T-cell receptor genes recently sequenced. (CS)