STRUCTURE OF CYTOTOXIC T LYMPHOCYTE GLYCOPROTEIN T145

细胞毒性T淋巴细胞是T淋巴细胞的一个亚群，它介导 机体对病毒感染或组织的免疫反应 异形文字。簇毛豌豆种子中的凝集素是 相当感兴趣，因为一种未经鉴定的凝集素制剂具有 已有报道可与小鼠的一种糖蛋白(T145)特异性结合 细胞毒性T淋巴细胞。在前期工作中，三种凝集素已经被 从簇毛花籽中提纯而成。这些凝集素是四聚体，是 由两个不同的亚基组成，具有不同的碳水化合物结合 具体细节。与B4凝集素的结合，这是这些凝集素中的主要凝集素 种子，克隆的细胞毒性T淋巴细胞数量显著增加 过度结合静息的脾细胞和胸腺细胞。在此应用程序中， 细胞毒T淋巴细胞克隆株上的糖蛋白(S)及其免疫原性 这种糖蛋白的低聚糖部分，它能与这种糖蛋白相互作用 用簇毛杆菌B4对凝集素进行了详细的研究。为了做到这一点，一辆V。 将制备簇毛霉菌B4凝集素亲和吸附剂并对其进行表征。 代谢性标记的细胞毒性T淋巴细胞上的糖蛋白 用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和荧光法对该吸附剂的结合力进行鉴定。 糖肽也将从这些细胞毒性T淋巴细胞和 簇毛芽孢杆菌B4凝集素-琼脂糖亲和层析分离 纵队。低聚糖单元将从标记的 与亲和柱相互作用的糖肽及其初级 结构已确定。一旦这种寡糖结构被阐明， 它产生的生物合成机制(S)将被研究。 最后，将抗血清提高到细胞毒性T细胞上的糖蛋白(S)。 与簇毛弧菌B4凝集素亲和吸附剂结合的淋巴细胞。 静息小鼠脾细胞和小鼠脾细胞上的免疫相似糖蛋白 胸腺细胞将通过免疫沉淀法进行鉴定 这些细胞中的糖蛋白。如果一种免疫相似的糖蛋白 在这些细胞上，蛋白质之间的结构相似之处 部分糖蛋白将用胰蛋白酶多肽进行表征。 映射。这些研究将提出一种机制，即糖蛋白 具有一种新的寡糖结构，在 细胞毒性T淋巴细胞的分化，最终将导致 更好地了解细胞表面糖蛋白和 他们在T淋巴细胞上的寡糖单位。

Cytotoxic T lymphocytes are a subpopulation of T lymphocytes which mediate the immune response of an organism to viral infection or tissue allographs. the lectins from the seeds of Vicia villosa are of considerable interest because an uncharacterized lectin preparation has been reported to bind specifically to a glycoprotein (T145) on mouse cytotoxic T lymphocytes. In preliminary work, three lectins have been purified from V. villosa seeds. These lectins are tetrameric and are composed of two different subunits with distinct carbohydrate binding specificities. Binding of B4 lectin, the predominant lectin in these seeds, to cloned lines of cytotoxic T lymphocytes is considerably increased over binding to resting splenocytes and thymocytes. In this application, the glycoprotein(s) on a clonel line of cytotoxic T lymphocytes and the oligosaccharide protion of this glycoprotein which specifically interacts with V. villosa B4 lectin will be studied in detail. To do this, a V. villosa B4 lectin affinity adsorbent will be prepared and characterized. The glycoproteins on metabolically-labeled cytotoxic T lymphocytes which bind to this adsorbent will be identified by SDS-PAGE and fluorography. Glycopeptides will also be prepared from these cytotoxic T lymphocytes and fractionated by affinity chromatography on the V. villosa B4 lectin-agarose column. Oligosaccharide units will be released from the labeled glycopeptides which interact with the affinity column and their primary structure determined. Once this oligosaccharide structure is elucidated, the biosythetic mechanism(s) by which it arises will be investigated. Finally, an antisera will be raised to the glycoprotein(s) on cytotoxic T lymphocytes which binds to the V. villosa B4 lectin affinity adsorbent. Immunologically similar glycoproteins on resting mouse splenocytes and thymocytes will be identified by immunoprecipitation of solubilized glycoproteins from these cells. If an immunologically similar glycoprotein is present on these cells, structural similarities between the protein portions of the glycoproteins will be characterized by tryptic peptide mapping. These studies will suggest a mechanism by which a glycoprotein with a new oligosaccharide structure is expressed during the differentiation of cytotoxic T lymphocytes and will eventually lead to a better understanding of the function of cell surface glycoproteins and their oligosacchride units on T lymphocytes.