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Direct transfection and activation of human cutaneous dendritic cells

Gene Therapy volume 8, pages 608–617 (2001)Cite this article

Abstract

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-1 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.

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Acknowledgements

We want to thank S Alber and C Almonte for their technical help in image acquisition, F Shagas and AC Bursick for their technical assistance with TEM techniques and the personnel from Magee Women's Hospital Labor and Delivery Department and Plastic Surgery Department for providing human skin samples. This work was supported by grants R21AI 469701 and PO1 AI 43664 from National Institutes of Health (LDF). Adriana T Larregina is a fellow of the Dermatology Foundation Research Career Development Program.

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Authors and Affiliations

  1. Department of Dermatology, University of Pittsburgh, School of Medicine, Pittsburgh, PA, USA

    A T Larregina, G Erdos, L A Spencer & L D Falo Jr

  2. University of Pittsburgh Cancer Institute, University of Pittsburgh, School of Medicine, Pittsburgh, PA, USA

    A T Larregina, S C Watkins, G Erdos, L A Spencer & L D Falo Jr

  3. Department of Cell Biology and Physiology, University of Pittsburgh, School of Medicine, Pittsburgh, PA, USA

    S C Watkins & D Beer Stolz

  4. Department of Surgery, Molecular Biology and Biochemistry and Pathology, University of Pittsburgh, School of Medicine, Pittsburgh, PA, USA

    W J Storkus

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  1. A T Larregina
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  3. G Erdos
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  4. L A Spencer
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  6. D Beer Stolz
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  7. L D Falo Jr
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Larregina, A., Watkins, S., Erdos, G. et al. Direct transfection and activation of human cutaneous dendritic cells. Gene Ther 8, 608–617 (2001). https://doi.org/10.1038/sj.gt.3301404

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