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: 2003504
• 负责人: ROBERT E. TIGELAAR
• 金额: $ 28.96万
• 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/2003504
• 关键词: CD antigens; T lymphocyte; cell adhesion molecules; cell differentiation; cellular immunity; clone cells; dendritic cells; fusion gene; gene expression; genetically modified animals; interleukin 2; interleukin 7; laboratory mouse; lymphocyte proliferation; 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.

该提案的目标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） 生长在所有正常小鼠的皮肤上，并了解它们的 与正常和患病人类皮肤中存在的 T 细胞的关系。 DETC 的独特特征（例如 V-gamma5/V-delta1+ 的同质性） T 细胞受体 (TCR)，成年小鼠对皮肤的明显限制， 以及识别受应激的表皮细胞上表达的配体的能力）， 导致了这样的假设：它们在皮肤中发挥着独特的作用 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、消除 改变/应激细胞，下调介导的反应 传统的 TCRα-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 的此类潜在相关生物学作用 通过开发同源的皮肤免疫和/或免疫病理学 选择性缺乏 V-gamma5+ DETC 的重组小鼠，然后进行测试 这种 DETC 缺陷小鼠在几个模型系统中，包括诱导和 皮肤肿瘤的生长、过敏性接触性皮炎和实验性 皮肤自身免疫性疾病。 2）影响因素的定义 DETC 在皮肤中的定位/增殖。 策略包括 尝试用 DETC 样重建 DETC 缺陷小鼠的皮肤 来自新生肝脏的细胞，以及生长期毛发作用的研究 通过分析 DETC 密度/异质性来了解这些过程中的卵泡 各种毛发突变小鼠的皮肤并通过荧光素的定位 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 分析广泛的正常和转化细胞/系 各种组织具有刺激 DETC 增殖的能力 体外分泌IL-2，使用α-热休克蛋白抗体或抗体 DETC 表达的各种粘附/辅助分子（例如 CD45、CD8、 LFA-1，CD28）尝试阻止此类反应，并准备 针对 DETC V-gamma5/V-delta1+ TCR 配体的单克隆抗体 应激表皮细胞以纯化相关刺激物 分子。 4）DETC功能异质性分析。 策略包括检查 DETC 中辅助分子的表达 不同的激活状态（新鲜分离与维持的细胞系/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 检查新鲜分离的淋巴因子和细胞毒性特征 从正常和 IL-2 缺陷小鼠中培养的 DETC（同源重组体） 或HSV-胸苷激酶转基因）与IL-2等细胞因子一起培养， IL-4或IL-7。此类研究应能增进对 γ-δ 的了解 生理和病理条件下细胞/皮肤的相互作用 老鼠和人。