STRUCTURAL BASIS OF CD59 AND CD58 SIGNALING TO T-CELLS

描述(改编自调查人员的摘要)：互动 T淋巴细胞和抗原提呈细胞之间的黏附 这些分子也被认为是细胞激活的调节器。 最近，研究人员表明，CD59抗原是第二种 除了公认的相互作用外，T细胞上CD2的配体 CD58(LFA-3)。这些细胞表面抗原之间的相互作用 在免疫调节中起着非常重要的作用，很可能形成第一个 理解黏附分子相互作用的范例。CD59是 也是补体膜形成的重要调节因素 攻击综合体。对其中重要因素的定义 相互作用最初将通过分子研究来完成。站点- 将对编码CD59和CD59的cDNA进行定向突变 和CD58抗原。突变株将使用单克隆进行筛选。 抗体、黏附试验和补体功能 CD59。随后，这些抗原的免疫原性将被 将其导入几种抗原提呈源后进行检查 细胞。重组蛋白将从CHO转染体或 杆状病毒表达系统并用于检测这三种病毒 空间结构。这些抗原以及MHC II类抗原 可能与内皮细胞的免疫原性有关。这个 转导CD58、CD59的猪内皮细胞的免疫原性 和/或人类MHC II类药物将在体外进行初步评估。这些 转基因细胞也将被用来确定猪 细胞有可能成为人体异种移植的潜在来源。 一种模型小鼠系统，其中SCID小鼠的免疫系统已经 用人类细胞重建的将被使用。免疫原性 修饰后的猪内皮细胞的性质将在 合成血管网络入门。这种方法还应该 提供对免疫介导的损伤的洞察。

DESCRIPTION (Adapted from the investigator's abstract):Interactions between T lymphocytes and antigen presenting cells involve adhesion molecules that are also implicated as regulators of cell activation. Recently, the investigators have shown that the CD59 antigen is a second ligand for CD2 on T cells in addition to the well recognized interaction with CD58 (LFA-3). The interactions among these cell surface antigens are very central to immune regulation and will likely form the first paradigm for understanding adhesion molecule interactions. The CD59 is also an important regulator of the formation of the complement membrane attack complex. Definition of the significant factors in this interaction will be accomplished initially with molecular studies. Site- directed mutagenesis will be performed on cDNAs encoding both the CD59 and CD58 antigens. The mutants will be screened using monoclonal antibodies, adhesion assays and for complement function in the case of CD59. Subsequently, the immunogenicity of these antigens will be examined after transfection into several sources of antigen presenting cells. Recombinant protein will be generated from CHO transfectants or baculovirus expression systems and used for determination of the three dimensional structure. These antigens as well as MHC class II antigens may contribute to the immunogenicity of endothelial cells. The immunogenicity of porcine endothelial cells transfected with CD58, CD59 and/or human MHC class II will be initially evaluated in vitro. These transfected cells will also be utilized to determine whether porcine cells could be a potential source for xenotransplantation into human. A model murine system in which the immune system of SCID mice has been reconstructed with human cells will be employed. The immunogenic properties of modified porcine endothelial cells will be evlauated after introduction in synthetic vascular networks. This approach should also provide insight into immune-mediated injury.