IMMORTALIZED HUMAN LANGERHANS CELLS

朗格汉斯细胞（Langerhans cell，LC）是皮肤的主要抗原提呈细胞， 在许多表皮现象中起关键作用， 皮炎和宿主防御机制对皮肤癌和 细菌、真菌和病毒感染。然而，目前的调查 由于以下原因，LC生物学和功能受到限制： 在培养物中不增殖和2）在标准培养条件下，LC 呈现出活化或引发的表型，就像抗原已经被呈递 并进行处理。因此，研究LC在其基础，未引发 国家严重受损。此外，将LC纳入 体外表皮组织模型，这将允许研究 从组织的角度来看，皮肤的免疫反应， 是可能的。为了克服这些问题，该项目将 开发永生化的人源性LC细胞系。 第一阶段将集中于建立和定性 永生化LC细胞系。正常LC将从人中收获 表皮并使用3种不同的方法永生化。所得的细胞 线将在生长速率、形态、表面 表型和超微结构。此外，细胞系的能力 刺激同种异体T细胞的能力将被确定。在第二阶段， 这些细胞系在研究接触性过敏性皮炎中的应用， 趋化因子受体和LC迁移将被证明， 最佳的细胞系将被纳入一个三维的 表皮皮肤模型 拟议的商业应用： 拟议的研究将产生稳定的LC线， 让我们更好地了解皮肤的免疫反应。 生产的LC线将为研究人员提供一种方法， LC在其天然、未引发状态下的生物学和功能可以是 研究了最后，研究人员将获得一个有价值的免疫工具， 研究和开发针对皮肤癌和细菌的疗法， 真菌和病毒感染，包括艾滋病毒。

As the main antigen-presenting cell of the skin, Langerhans cells (LC) play a key role in many epidermal phenomena such as contact allergic dermatitis and in host defense mechanisms against skin cancer and bacterial, fungal, and viral infections. However, current investigations into LC biology and function are limited due to the following: l) LC do not proliferate in culture and 2) under standard culture conditions, LC take on an activated or primed phenotype as if antigen had been presented and processed. Thus, the ability to study LC in their basal, unprimed state is severely compromised. In addition, the incorporation of LC into in vitro epidermal tissue models, which would allow the study of the immunological reactions of the skin from a tissue perspective, has not been possible. In order to overcome these problems, this project will develop immortalized, human-derived LC lines. Phase I will concentrate on the establishment and characterization of immortalized LC cell lines. Normal LC will be harvested from human epidermis and immortalized using 3 different methods. The resultant cell lines will be characterized in terms of growth rates, morphology, surface phenotype, and ultrastructure. In addition, the ability of the cell lines to stimulate allogeneic T-cells will be determined. During Phase II, the utility of these cell lines to study contact allergic dermatitis, chemokine receptors, and LC migration will be demonstrated and the optimal cell lines will be incorporated into a three-dimensional epidermal skin model. PROPOSED COMMERCIAL APPLICATIONS: The proposed research will produce stable LC lines which will ultimately allow a greater understanding of the immunological reactions of the skin. The LC lines produced will provide researchers the means by which the biology and function of LC in their native, unprimed state can be studied. Finally, researchers will gain a valuable immunological tool in studying and developing therapies against skin cancer and bacterial, fungal, and viral infections including HIV.