ROLE OF FUCOSYLTRANSFERASE IN ANTIVIRAL CYTOTOXICITY

描述（改编自研究者摘要）：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，它们在体内的分化仅被部分理解。我们有 发现一种新的APC-CD 8 T细胞相互作用通过α（1，3）岩藻糖基化的 P-选择素糖蛋白-1（PSGL-1）是产生效应CTL所必需的。在其 缺乏（1，3）岩藻糖基转移酶的牛痘病毒感染小鼠 IV+VII（FT-/-）在产生效应CTL方面是缺陷的，尽管它们产生 活化的CD 8 T细胞能够病毒特异性增殖。目标1： 将FT-/-小鼠培育成淋巴细胞性脉络丛脑膜炎转基因（Tg）小鼠 （LCMV）糖蛋白TCR。这一系统将使我们能够研究金融时报的作用， LCMV攻击后的抗原特异性CTL应答。我们将确定FT是否 在过继性转移实验中，APC或CD 8 T细胞上需要。白细胞 II型粘附缺陷（LADII），一种由岩藻糖缺陷引起的人类疾病 代谢使我们能够研究岩藻糖基化PSGL-1在人体中的作用。 我们还培育出了T细胞表达水母基因的转基因小鼠 维多利亚水母衍生的绿色荧光蛋白（GFP-Tg）。当这些老鼠 免疫后，效应CTL失去GFP表达，而幼稚和 活化的前效应T细胞继续表达GFP。因此，GFP-Tg小鼠 提供了一个强有力的工具，以区分效应子和前效应子 CTL.使用这个系统，我们已经确定了几个以前没有认识到的 在效应分化过程中发生的分子变化。因此，为了研究 缺乏FT的分子机制是有缺陷的，在目标2中，我们 将FT-/-小鼠培育成GFP-Tg小鼠，并分析它们的抗原活化的CD 8 T 细胞与FT+/+ CD 8 T细胞的分子差异。 由于选择素-配体缺陷型FT-/-小鼠，而不是选择素缺陷型小鼠， 对于CTL产生缺陷，非选择素受体可能存在于 岩藻糖基化PSGL-1。事实上，我们已经确定了这样一个反受体， 使用可溶性重组PSGL-1四聚体活化T细胞。在目标3中，我们 将分离、克隆和鉴定该分子。

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.