CHEMOKINE REGULATION OF NKT CELLS, AND ACAID

NKT 细胞的趋化因子调节和 ACAID

• 批准号: 6525048
• 负责人: Joan Stein-Streilein
• 金额: $ 40.68万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6525048
• 关键词: chemokine; cytokine receptors; cytotoxic T lymphocyte; delayed hypersensitivity; eye; genetically modified animals; immune tolerance /unresponsiveness; laboratory mouse; leukocyte activation /transformation; macrophage; monocyte; natural killer cells

Chemokines are associated with a variety of inflammatory and autoimmune diseases for their role in recruitment of cells to the sites of immune induction and inflammation. However, few studies have explored a role for chemokines in peripheral tolerance induction and no studies are reported for specific chemokines for NKT cells. This research plan will explore the role of specific chemokines in negative regulation of an immune inflammatory response and specifically in the recruitment of NKT cells to the site of tolerance induction. Anterior Chamber Associated Immune Deviation (ACAID) is associated with the selective negative regulation of delayed type hypersensitivity (DTH) after the introduction of antigen into the eye. An accumulation of splenic NKT cells is absolutely required for the development of ACAID. Preliminary data show that MIP-2 is associated with NKT cell recruitment. We will use molecular, cellular and morphological techniques to identify and define the role of the chemokines in the tolerance inducing process. Aim One will define the chemokine profile by RNase Protection Assay and multi-color flow cytometry. The second aim will use classic chemotaxis assays to define the migratory response of NKT cells to ACAID associated chemokines, along with molecular techniques to confirm the expression of chemokine receptors on these cells. The third aim will explore the mechanisms of chemokine-NKT cell interaction during ACAID induction by reconstituting NKT cell knockout mice with NKT cells from chemokine receptor knockout mice. A local adoptive transfer assay will measure if regulator T cells were generated. Finally, the fourth aim will explore the hypothesis that the cells recruited to the spleen for the induction of ACAID must necessarily form interactive cell clusters. After identifying such clusters and cells within, we will modulate chemokines and potential adhesion molecules and evaluate the outcome by quality and quantity of clusters. NKT cell defects and/or deficiency is associated with a variety of autoimmune disorders in both mice and humans (lupus, type 1 diabetes, systemic scleroderma). Thus, the mechanisms of NKT cell mediated induction of T regulatory cells might function in the maintenance of self tolerance in a variety of organs and tissues in addition to immune privileged sites such as the eye. The novel postulates about chemokines, innate cells and peripheral tolerance presented in this proposal, represent a heretofore totally unexplored area of research.

趋化因子与多种炎症和自身免疫性疾病有关，因为它们在将细胞募集到免疫诱导和炎症部位中的作用。然而，很少有研究探索趋化因子在外周耐受性诱导中的作用，也没有关于NKT细胞特异性趋化因子的研究报道。该研究计划将探索特异性趋化因子在免疫炎症反应负调控中的作用，特别是在NKT细胞募集到耐受诱导部位中的作用。前房相关免疫偏差（ACAID）与抗原进入眼后延迟型超敏反应（DTH）的选择性负调控有关。脾NKT细胞的积累对于ACAID的发展是绝对必要的。初步数据显示MIP-2与NKT细胞募集有关。我们将使用分子、细胞和形态学技术来识别和定义趋化因子在诱导耐受性过程中的作用。目的一将通过rna酶保护实验和多色流式细胞术确定趋化因子的谱。第二个目标将使用经典的趋化性分析来确定NKT细胞对ACAID相关趋化因子的迁移反应，并使用分子技术来确认趋化因子受体在这些细胞上的表达。第三个目标是通过用趋化因子受体敲除小鼠的NKT细胞重组NKT细胞敲除小鼠，探索ACAID诱导过程中趋化因子-NKT细胞相互作用的机制。局部过继转移试验将测量是否产生调节T细胞。最后，第四个目标将探讨一个假设，即为诱导ACAID而招募到脾脏的细胞必须形成相互作用的细胞团。在确定这些簇和细胞后，我们将调节趋化因子和潜在的粘附分子，并通过簇的质量和数量评估结果。NKT细胞缺陷和/或缺乏与小鼠和人类的多种自身免疫性疾病（狼疮、1型糖尿病、系统性硬皮病）有关。因此，NKT细胞介导的诱导T调节细胞的机制可能在多种器官和组织中发挥作用，除了免疫特权部位（如眼睛）。本提案中提出的关于趋化因子、先天细胞和外周耐受性的新假设，代表了一个迄今为止完全未探索的研究领域。