ROLE OF FUCOSYLTRANSFERASE IN ANTIVIRAL CYTOTOXICITY

岩藻糖基转移酶在抗病毒细胞毒性中的作用

• 批准号: 6707535
• 负责人: MANJUNATH NARASIMHA SWAMY
• 金额: $ 42.57万
• 依托单位: IMMUNE DISEASE INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-15 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6707535
• 关键词: CD95 molecule; T cell receptor; antigen presentation; antigen presenting cell; cellular immunity; clinical research; cytotoxic T lymphocyte; enzyme activity; genetically modified animals; green fluorescent proteins; hexosyltransferase; human subject; immunodeficiency; interferon gamma; interleukin 2; interleukin 4; laboratory mouse; leukocyte activation /transformation; lymphocytic choriomeningitis virus; passive immunization; pore forming protein; selectins; tumor necrosis factor alpha

DESCRIPTION (adapted from investigator's abstract): Cytotoxic T lymphocytes (CTL) are key immune effector cells in controlling viral infections and tumors. However, their differentiation in vivo is only partially understood. We have found that a novel APC-CD8 T cell interaction through an a(1,3) fucosylated P-selectin glycoprotein-1 (PSGL-1) is required to generate effector CTL. In its absence, Vaccinia virus-infected mice deficient in a(1,3) fucosyl transferase IV+VII (FT-/-) are deficient in generating effector CTL, although they generate activated CD8 T cells capable of viral-specific proliferation. In Aim 1, we will breed FT-/- mice to mice transgenic (Tg) to lymphocytic choriomeningitis (LCMV) glycoprotein TCR. This system will enable us to study the role of FT in antigen-specific CTL response after LCMV challenge. We will determine if FT is required on APC or on CD8 T cells in adoptive transfer experiments. Leukocyte adhesion deficiency type II (LADII), a human disease due to defective fucose metabolism allows us to study the role of fucosylated PSGL-1 in humans. We have also produced transgenic mice whose T cells express the jellyfish Victoria acquoria-derived green fluorescent protein (GFP-Tg). When these mice are immunized, the effector CTL lose GFP expression, whereas naive and activated pre-effector T cells continue to express GFP. Thus, GFP-Tg mice provide a powerful tool to separate differentiated effector from pre-effector CTL. Using this system, we have identified several previously unrecognized molecular changes that occur during effector differentiation. Thus, to study the molecular mechanisms that are defective in the absence of FT, in aim 2 we will breed FT-/- mice to GFP-Tg mice and analyze their antigen activated CD8 T cells for molecular differences from FT+/+ CD8 T cells. Since selectin-ligand deficient FT-/- mice, but not selectin-deficient mice are defective for CTL generation, a non-selectin receptor may exist for the fucosylated PSGL-1. Indeed, we have identified such a counter-receptor on activated T cells using soluble recombinant PSGL-1 tetramers. Thus in Aim 3, we will isolate, clone and characterize the molecule.

描述(摘自研究人员摘要)：细胞毒性T淋巴细胞 CTL是控制病毒感染和肿瘤的关键免疫效应细胞。 然而，对它们在体内的分化只有部分了解。我们有 发现一种新的APC-CD8 T细胞通过a(1，3)岩藻糖基化相互作用 P-选择素糖蛋白-1(PSGL-1)是产生效应性CTL所必需的。在ITS中 缺乏(1，3)岩藻糖基转移酶的痘苗病毒感染小鼠 IV+VII(FT-/-)在产生效应器CTL方面存在缺陷，尽管它们产生 激活的CD8 T细胞能够进行病毒特异性增殖。在目标1中，我们 将培育转(TG)淋巴细胞性脉络膜脑膜炎的FT-/-小鼠 (LCMV)糖蛋白TCR。这一系统将使我们能够研究金融时报在 LCMV攻击后的抗原特异性CTL反应。我们将确定FT是否 在过继转移实验中，APC或CD8 T细胞上需要。白细胞 黏附缺陷II型(LADII)，一种由岩藻糖缺陷引起的人类疾病 新陈代谢使我们能够研究岩藻糖基化PSGL-1在人类中的作用。 我们还培育出了T细胞表达水母的转基因小鼠 维多利亚金雀花衍生绿色荧光蛋白(GFP-TG)。当这些老鼠 免疫后，效应器CTL失去GFP表达，而幼稚和 激活的前效应子T细胞继续表达GFP。因此，GFP-TG小鼠 提供强大的工具来区分区分的效应器和前效应器 CTL.使用这个系统，我们已经确定了几个以前没有被识别的 在效应器分化过程中发生的分子变化。因此，要研究 在目标2中，我们研究了在没有FT的情况下有缺陷的分子机制 将FT-/-小鼠培育成GFP-TG小鼠并分析其抗原活化的CD8 T细胞 T细胞与FT+/+CD8T细胞的分子差异。 因为选择素配体缺陷FT-/-小鼠，而不是选择素缺陷小鼠 CTL生成有缺陷，可能存在非选择素受体 岩藻糖基化PSGL-1。事实上，我们已经确定了这样一种受体拮抗剂 用可溶性重组PSGL-1四聚体激活T细胞。因此，在目标3中，我们 将分离、克隆和鉴定该分子。