Direct transfection and activation of human cutaneous dendritic cells
Direct transfection and activation of human cutaneous dendritic cells
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DOI：
10.1038/sj.gt.3301404
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发表时间：
2001-04-01
期刊：
影响因子：
5.1
通讯作者：
Falo, LD
中科院分区：
文献类型：
--
作者：
Larregina, AT;Watkins, SC;Falo, LD
基因治疗技术可以成为诱导和控制免疫反应的重要工具。抗原递送是疫苗设计中的一个关键挑战，基于dna的免疫为递送编码的转基因蛋白抗原提供了一种有吸引力的方法。在本研究中，我们使用基因枪转染人类皮肤器官培养物，目的是在皮肤树突状细胞中表达转基因抗原。我们的研究表明，当输送到人体皮肤时，即使使用高氦输送压力，也主要在表皮中观察到金颗粒。我们证明了居住在基底表皮的朗格汉斯细胞可以被转染，并且生物基因传递足以刺激皮肤树突状细胞的激活和迁移。对从转染皮肤中迁移的树突状细胞进行RT-PCR分析，发现存在转基因mRNA，表明直接转染了皮肤树突状细胞。重要的是，转染的表皮朗格汉斯细胞可以有效地将来自转基因黑色素瘤抗原mart -1的肽呈递到mart -1特异性CTL。综上所述，我们的结果证明了原位转导的人朗格汉斯细胞的直接转染、激活和抗原特异性刺激功能。
Gene therapy techniques can be important tools for the induction and control of immune responses. Antigen delivery is a critical challenge in vaccine design, and DNA-based immunization offers an attractive method to deliver encoded transgenic protein antigens. In the present study, we used a gene gun to transfect human skin organ cultures with a particular goal of expressing transgenic antigens in resident cutaneous dendritic cells. Our studies demonstrate that when delivered to human skin, gold particles are observed primarily in the epidermis, even when high helium delivery pressures are used. We demonstrate that Langerhans cells resident in the basal epidermis can be transfected, and that biolistic gene delivery is sufficient to stimulate the activation and migration of skin dendritic cells. RT-PCR analysis of dendritic cells, which have migrated from transfected skin, demonstrates the presence of transgenic mRNA, indicating direct transfection of cutaneous dendritic cells. Importantly, transfected epidermal Langerhans cells can efficiently present a peptide derived from the transgenic melanoma antigen MART-I to a MART-1-specific CTL. Taken together, our results demonstrate direct transfection, activation, and antigen-specific stimulatory function of in situ transduced human Langerhans cells.