BIOLOGY OF THE THY-1 ANTIGEN-BEARING EPIDERMAL CELLS

携带 THY-1 抗原的表皮细胞的生物学

• 批准号: 2079011
• 负责人: AKIRA TAKASHIMA
• 金额: $ 22.31万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-01 至 1997-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2079011
• 关键词: Langerhans' cell; T lymphocyte; cell transformation; cellular immunity; cytokine; dendritic cells; genetically modified animals; interferon gamma; keratinocyte; laboratory mouse; skin neoplasms; tissue /cell culture; transforming growth factors

The objective of this proposal is to characterize mechanism by which keratinocytes (KC) cooperate with gamma delta T cells to form an immunological defense at the environmental interface. Working hypotheses are that: 1) KC and resident gamma delta T cells play critical roles in maintaining the immunological integrity of skin, 2) certain functions of KC and gamma delta T cells are regulated by locally produced cytokines, and 3) environmental stimuli disrupt this equilibrium by altering these cytokine profiles. Dendritic Epidermal T Cells (DETC), one member of the family of resident epithelial gamma delta T cells in mice, will be studied as a prototypic cutaneous gamma delta T cell. IL-7 and transforming growth factor-Beta (TGFBeta) will be studied as cytokines produced by KC that regulate T cell function, and gamma-interferon (gammaIFN) will be studied as a cytokine produced by T cells that modulates KC function. These hypotheses are derived from observations that: 1) DETC growth is promoted by IL-7 and inhibited by TGFbeta, 2) external stimuli modulate cytokine production in KC (e.g., UVB-induced downregulation of IL-7 MRNA expression and gamma IFN-induced expression of a truncated (2.6 kb) IL-7 transcript, 3) DETC kill skin tumor targets and downregulated alpha/beta T cell-mediate immunity, and 4) gamma delta T cells infiltrate and play effector roles in several human skin disorders. Specific aims are: 1) To determine the physiological roles of IL-7 and TGFbeta in modulating DETC function. Local IL-7 and TGFBeta concentrations will be altered experimentally [cytokine or mAb injection, IL-7 transgenic mice] and DETC in test will be examined for maturation during the neonatal period, modulation of their cell densities, and killing activities. 2) To study IL-7 and TGFBeta gene regulation in KC: a) modulation of IL-7 or TGFBeta production in KC by environmental and cytokine stimuli [Northern blotting, bioassays]; and b) roles of early immediate genes in this regulation [antisense nucleotide blocking]. 3) To identify the function of a gammaIFN-inducible 2.6 kb IL-7 transcript [cDNA cloning and sequencing, mammalian cell expression]. 4) To characterize in vivo functions of DETC. DETC densities will be manipulated experimentally and skin response to alpha/Beta T cell- mediated attack will be examined. 5) To define mechanisms for the cytotoxic recognition of skin tumors by DETC: a) the role played by 2 B4, a novel molecule that mediates non-MHC-restricted target recognition, including b) regulation of 2B4 expression. The proposed studies should lead to a better understanding of cytokine networks in epidermis, the biology of cutaneous gamma delta T cells, and the pathophysiology of T cell-mediated and neoplastic skin disorders in humans.

该提议的目的是表征机制 角质形成细胞（KC）与伽马三角细胞合作以形成 环境界面的免疫防御。 工作假设 是：1）KC和常驻伽马三角洲T细胞在 维持皮肤的免疫学完整性，2） KC和Gamma Delta T细胞由局部产生的细胞因子调节， 3）环境刺激通过改变这些均衡来破坏这种平衡 细胞因子谱。 树突状表皮T细胞（DETC），其中一个成员 小鼠中常驻上皮伽马三角细胞的家族将是 研究为原型皮肤伽马三角细胞。 IL-7和 转化生长因子β（TGFBETA）将作为细胞因子研究 由调节T细胞功能的KC和伽马 - 互换的KC产生 （γFN）将研究作为由T细胞产生的细胞因子 调节KC功能。 这些假设来自观察 那就是：1）IL-7促进了开特的生长，并被TGFBETA抑制，2） 外部刺激调节KC中的细胞因子产生（例如，UVB诱导的 IL-7 mRNA表达和伽马IFN诱导表达的下调 截短的（2.6 kb）IL-7转录本，3）detc杀死皮肤肿瘤靶标 并下调α/βT细胞 - 介质免疫力，以及4）伽马三角洲 T细胞浸润和扮演效应子在几种人皮肤中的作用 疾病。 具体目的是：1）确定生理角色 在调节发电函数中的IL-7和TGFBETA的of。 本地IL-7和TGFBETA 浓度将通过实验[细胞因子或mAb注射， 将检查测试中的IL-7转基因小鼠]和DETC的成熟 在新生儿时期，调节其细胞密度和 杀人活动。 2）研究KC中的IL-7和TGFBETA基因调节： a）通过环境和 细胞因子刺激[北印迹，生物测定]； b）早期的角色 该调节中的直接基因[反义核苷酸阻断]。 3） 确定γFN诱导的2.6 kb IL-7转录本的功能 [cDNA克隆和测序，哺乳动物细胞表达]。 4）到 表征destc的体内功能。 detc密度将是 通过实验和对α/βT细胞的皮肤反应进行操纵 - 将检查中介攻击。 5）定义机制 DETC对皮肤肿瘤的细胞毒性识别：a）2 B4的作用， 一种介导非MHC限制靶标识别的新型分子， 包括b）2B4表达的调节。 拟议的研究应 使对表皮中的细胞因子网络有更好的了解， 皮肤伽马三角T细胞的生物学和T的病理生理学 细胞介导的和肿瘤的皮肤疾病。