Lymphocyte Migration in Development of Type 1 Diabetes

描述（由申请人提供）： 目标 1型糖尿病（T1 DM）是一种T细胞介导的自身免疫性疾病。在自身免疫应答的早期阶段，初始自身反应性T细胞从血液迁移到胰腺淋巴结中，在那里它们遇到β细胞抗原;这些T细胞的记忆/效应子细胞然后从血液迁移到胰岛中以引发导致β细胞破坏的炎症。淋巴细胞从血液迁移到组织中需要组织选择性的多步级联反应，具有顺序的淋巴细胞/内皮细胞粘附和活化步骤。我们的目标是确定在炎症早期胰腺淋巴结和胰岛中表达的内皮粘附和活化分子（如趋化因子），以确定这些分子在淋巴细胞募集到这些部位和启动自身免疫反应中的作用，并确定哪些分子是预防T1 DM的治疗靶点。 研究计划和方法 我们将使用组织切片免疫组织学，悬浮液免疫荧光染色流式细胞术，激光捕获显微切割（LCM）与RT-PCR，以确定哪些粘附分子，趋化因子和趋化因子受体存在于胰腺淋巴结（目的1）和胰岛（目的2）的非肥胖糖尿病（NOD）小鼠在自身免疫过程的早期阶段。这些分子在幼稚自身反应性T细胞迁移到胰腺淋巴结（Aim 1）和产生迁移到胰岛的记忆/效应T细胞（Aim 2）中的生理作用将使用体内淋巴细胞迁移测定来定义。在目标3中，我们将与Hugh McDevitt博士密切合作，以确定TNF α和抗TNF α对淋巴细胞迁移途径和自身免疫或耐受发展的影响。

DESCRIPTION (provided by applicant): OBJECTIVES Type 1 diabetes (T1DM) is a T cell mediated autoimmune disease. In the early stages of the autoimmune response, naive autoreactive T cells migrate from blood into pancreatic lymph nodes where they meet Beta cell antigens; memory/effector offspring of these T cells then migrate from blood into islets to initiate the inflammation that leads to Beta cell destruction. The migration of lymphocytes from blood into tissues requires tissue-selective multistep cascades with sequential lymphocyte/endothelial adhesion and activation steps. Our goals are to identify the endothelial adhesion and activating molecules (such as chemokines) that are expressed in pancreatic lymph nodes and islets in the early stages of inflammation, to define the roles of these molecules in lymphocyte recruitment to these sites and in the initiation of the autoimmune response, and to determine which molecules are therapeutic targets for the prevention of T1DM. RESEARCH PLANS AND METHODS We will use tissue section immunohistology, suspension immunofluorescence staining with flow cytometry, and laser capture microdissection (LCM) with RT-PCR to determine which adhesion molecules, chemokines, and chemokine receptors are present in pancreatic lymph nodes (Aim 1) and islets (Aim 2) of nonobese diabetic (NOD) mice in the early stages of the autoimmune process. The physiologic roles of these molecules in the migration of naive autoreactive T cells into pancreatic lymph nodes (Aim 1) and in the production of memory/effector T cells which migrate to islets (Aim 2) will be defined using in vivo lymphocyte migration assays. In Aim 3, we will work closely with Dr. Hugh McDevitt to determine the effects of TNFalpha and anti- TNFalpha on lymphocyte migration pathways and on the development of autoimmunity or tolerance.