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.

趋化因子在将细胞募集到免疫诱导和炎症部位方面发挥作用，因此与多种炎症和自身免疫性疾病相关。 然而，很少有研究探讨趋化因子在外周耐受诱导中的作用，并且没有报道针对 NKT 细胞的特定趋化因子的研究。 该研究计划将探讨特定趋化因子在免疫炎症反应的负调节中的作用，特别是在将 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）与将抗原引入眼后迟发型超敏反应（DTH）的选择性负调节相关。 ACAID 的发生绝对需要脾脏 NKT 细胞的积累。初步数据表明，MIP-2 与 NKT 细胞招募相关。 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耐受诱导过程中的作用。 Aim One 将通过 RNase 保护测定和多色流式细胞术定义趋化因子谱。 第二个目标将使用经典的趋化性测定来定义 NKT 细胞对 ACAID 相关趋化因子的迁移反应，并利用分子技术来确认这些细胞上趋化因子受体的表达。 第三个目标是通过用趋化因子受体敲除小鼠的 NKT 细胞重建 NKT 细胞敲除小鼠，探讨 ACAID 诱导过程中趋化因子与 NKT 细胞相互作用的机制。 局部过继转移测定将测量调节性 T 细胞是否产生。 最后，第四个目标将探讨以下假设：为诱导 ACAID 而募集到脾脏的细胞必须形成相互作用的细胞簇。 在识别出此类簇和其中的细胞后，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来评估结果。 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，除了眼睛等免疫特权部位外，NKT 细胞介导的 T 调节细胞诱导机制可能在多种器官和组织的自我耐受维持中发挥作用。 该提案中提出的关于趋化因子、先天细胞和外周耐受的新颖假设代表了迄今为止完全未经探索的研究领域。