IMMUNOBIOLOGY OF MURINE DENDRITIC EPIDERMAL T-CELLS

鼠树突状表皮 T 细胞的免疫生物学

• 批准号: 2063828
• 负责人: ROBERT E. TIGELAAR
• 金额: $ 25.91万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-01-01 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2063828
• 关键词: T lymphocyte; cell adhesion molecules; cell differentiation; cellular immunity; clone cells; dendritic cells; fusion gene; gene expression; genetically modified animals; interleukin 2; laboratory mouse; lymphocyte proliferation; lymphokines; monoclonal antibody; skin; stress proteins

The goals of this proposal are to understand the immunobiologic relevance o a unique subset of T cells (called dendritic epidermal T cells, or DETC) which populate the skin of all normal mice, and to understand their relationship(s) to the T cells present in normal and diseased human skin. The unique features of DETC (e.g., homogeneity of their V-gamma5/V-delta1+ T cell receptors (TCRs), apparent restriction in adult mice to the skin, and capacity to recognize a ligand expressed on stressed epidermal cells), have led to the hypothesis that they play distinctive roles in cutaneous immune surveillance and/or immunoregulation; postulated (but unproven) roles include early responses to intracellular infection, elimination of altered/stressed cells, down-regulation of responses mediated by conventional TCRalpha-beta cells, and perhaps even initiation of autoimmunity. The specific aims of this proposal include: 1) In vivo examination of such potentially relevant biologic roles of DETC in cutaneous immunity and/or immunopathology by developing homologous recombinant mice selectively deficient in V-gamma5+ DETC and then testing such DETC-deficient mice in several model systems, including induction and growth of skin tumors, allergic contact dermatitis, and experimental cutaneous autoimmune disease. 2) Definition of factors affecting the localization/proliferation of DETC in the skin. Strategies include attempts to reconstitute the skin of DETC-deficient mice with DETC-like cells from newborn liver, and studies of the role(s) of anagen hair follicles in these processes by analysis of DETC density/heterogeneity in the skin of various hair mutant mice and by localization of fluorescein- labeled fetal thymocytes. 3) Characterize the ligands which activate DETC and the molecules involved in such activation. Strategies include analysis of a broad range of normal and transformed cells/lines from various tissues for their abilities to stimulate DETC proliferation and IL-2 secretion in vitro, use of alpha-heat shock protein Abs or Abs to various adhesion/accessory molecules expressed by DETC (such as CD45, CD8, LFA-1, CD28) to try to block such responses, and preparation of a monoclonal Ab directed against the DETC V-gamma5/V-delta1+ TCR ligand on stressed epidermal cells in order to purify the relevant stimulatory molecule(s). 4) Analysis of the functional heterogeneity of DETC. Strategies include examining accessory molecule expression on DETC in different activation states (freshly isolated vs lines/clones maintained under distinct culture condition) and correlating these patterns with DETC proliferation, lymphokine secretion and cytotoxicity profiles, as well as examining the lymphokine and cytotoxicity profiles of freshly isolated and cultured DETC from normal and IL-2-deficient mice (homologous recombinants or HSV-thymidine kinase transgenics) cultured with such cytokines as IL-2, IL-4 or IL-7. Such studies should advance understanding of gamma-delta cell/skin interactions under physiologic and pathologic circumstances in mice and in man.

这项建议的目的是了解免疫生物学相关性， o独特的T细胞亚群（称为树突状表皮T细胞或DETC） 它们分布在所有正常小鼠的皮肤上， 与存在于正常和患病的人皮肤中的T细胞的关系。 DETC的独特功能（例如，其V-γ 5/V-δ 1+均匀性 T细胞受体（TCR），成年小鼠对皮肤的明显限制， 和识别在应激表皮细胞上表达的配体的能力）， 这导致了一种假设，即它们在皮肤组织中起着独特的作用， 免疫监视和/或免疫调节;假设（但未经证实） 作用包括对细胞内感染的早期反应， 改变/应激细胞，下调由 传统的TCR α-β细胞，甚至可能启动 自身免疫该提案的具体目标包括：1）体内 检查DETC在以下方面的潜在相关生物学作用： 皮肤免疫和/或免疫病理学， 选择性缺乏V-γ 5 + DETC的重组小鼠，然后测试 在几种模型系统中，包括诱导和 皮肤肿瘤生长、过敏性接触性皮炎和实验性 皮肤自身免疫病2)影响因素的定义 DETC在皮肤中的定位/增殖。 策略包括 试图用DETC样蛋白重建DETC缺陷小鼠的皮肤， 新生肝细胞，以及生长期毛发作用的研究 通过分析DETC密度/异质性， 各种毛发突变小鼠的皮肤，并通过荧光素- 标记的胎儿胸腺细胞。3)表征激活DETC的配体 以及参与这种激活的分子。 策略包括 对来自人的广泛的正常和转化细胞/系的分析 各种组织刺激DETC增殖的能力， 体外IL-2分泌，α-热休克蛋白Ab或Ab用于 由DETC表达的各种粘附/辅助分子（如CD 45，CD 8， LFA-1，CD 28）来尝试阻断这种反应，并制备了一种免疫抑制剂。 针对DETC V-gamma 5/V-delta 1 + TCR配体的单克隆抗体 强调表皮细胞，以纯化相关的刺激 分子。4)DETC功能异质性分析。 策略包括检查DETC上的辅助分子表达， 不同的激活状态（新鲜分离的vs维持的系/克隆 在不同的培养条件下），并将这些模式与DETC相关联 增殖、淋巴因子分泌和细胞毒性特征，以及 检查淋巴因子和细胞毒性概况的新鲜分离， 来自正常和IL-2缺陷小鼠的培养的DETC（同源重组体 或HSV-胸苷激酶转基因）与细胞因子如IL-2， IL-4或IL-7。这样的研究应该会促进对伽玛-δ的理解 在生理和病理情况下的细胞/皮肤相互作用， 小鼠和人类。