DENDRITIC CELL TARGETED CONDITIONAL GENE EXPRESSION MODEL SYSTEMS

树突状细胞靶向条件基因表达模型系统

• 批准号: 6100563
• 负责人: Kiyoshi Ariizumi
• 金额: $ 5.51万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6100563
• 关键词: Langerhans' cell; dendritic cells; disease /disorder model; gene expression; gene targeting; genetic regulatory element; genetically modified animals; laboratory mouse; leukocyte activation /transformation; lymphocyte proliferation; model design /development; skin; tetracyclines; tissue /cell culture

Dendritic cells (DC) and Langerhans cells (LC) are distinguished by their potent antigen presenting cell (APC) capacity for activating T- cells, particularly in 1/0 immune responses. Application of gene- targeted techniques to investigate the biology of these cells has been hampered by a lack of well-defined DC/LC-specific regulatory gene elements. We have identified a 5'-flanking region (5FR) of the DC/LC- specific dectin-2 gene that can induce mRNA synthesis in a DC/LC- specific manner. The goal of this pilot and feasibility study is to develop a novel mouse model in which dectin-2 5FR-driven gene expression can be conditionally manipulated so as to permit depletion of DC/LC locally or systemically. To define logistics and optimal requirements we will first establish transient DC/LC-targeted systems in vitro and in mouse skin using dectin-2 5FR, the luciferase gene, and the Tet-On system, in which exogenously administered doxycycline can induce gene expression. Subsequently, we will generate transgenic mice bearing DC/LC-targeted luciferase expression also conditioned by doxycycline. Finally, we will develop dectin-2 5FR-driven, doxycycline responsive transgenic mice that carry the Herpes simplex thymidine kinase gene, in which DC/LC can be depleted by treatment with ganciclovir locally or systemically. These DC/LC-targeted conditional gene expression model systems will be valuable tools for amplifying our understanding of DC/LC biology. Establishment of these model systems will open the door to several other genes that can be targeted for DC/LC-specific expression. Ultimately, our depletion studies may prove applicable to the management of patients with histiocytosis X, a potentially fatal disease of unrestrained LC proliferation for which no satisfactory treatment currently exists.

树突状细胞（DC）和Langerhans细胞（LC）通过 它们的有效抗原呈现细胞（APC）激活T- 细胞，特别是在1/0免疫反应中。基因的应用 研究这些细胞生物学的有针对性技术已经 由于缺乏定义明确的DC/LC特异性调节基因而阻碍 元素。我们已经确定了DC/LC-的5'乘区域（5fr） 可以在DC/LC-中诱导mRNA合成的特定Dectin-2基因 具体方式。该飞行员和可行性研究的目的是 开发一种新型的小鼠模型，其中Dectin-2 5fr驱动基因表达 可以有条件地操纵以允许DC/LC耗尽 本地或系统地。定义物流和最佳要求 我们将首先在体外建立瞬态DC/LC靶向系统 使用Dectin-2 5fr，荧光素酶基因和Tet-On中的小鼠皮肤中的皮肤 系统，外源给予强力霉素可以诱导基因 表达。随后，我们将产生转基因小鼠轴承 DC/LC靶向的荧光素酶表达也由强力霉素调节。 最后，我们将开发dectin-2 5fr驱动的强力霉素响应 携带疱疹单纯胸苷激酶基因的转基因小鼠在 哪个DC/LC可以通过在本地使用Ganciclovir处理或 全身。这些DC/LC靶向条件基因表达模型 系统将是放大我们对DC/LC的理解的宝贵工具 生物学。建立这些模型系统将为 可以针对DC/LC特异性表达的其他几个基因。 最终，我们的耗竭研究可能适用于管理层 组织细胞增多症X的患者，这是一种潜在的致命疾病 不受约束的LC增殖，没有令人满意的治疗 目前存在。