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Introduction of functional chimeric E/L-selectin by RNA electroporation to target dendritic cells from blood to lymph nodes

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Abstract

Background

Inefficient migration of dendritic cells (DC) to regional lymph nodes (LN) upon intracutaneous injection is a major obstacle for effective DC vaccination. Intravenous vaccination is unfavorable, because DC cannot migrate directly from the blood into LN.

Methods

To enable human monocyte-derived (mo)DC to enter LN directly from the blood, we manipulated them by RNA electroporation to express a human chimeric E/L-selectin (CD62E/CD62L) protein, which binds to peripheral node addressin expressed on high endothelial venules.

Results

Transfection efficiency exceeded 95%, and high E/L-selectin surface expression was detected for >48 h. E/L-selectin RNA-transfected DC displayed an identical mature DC phenotype as mock-transfected DC. Furthermore, E/L-selectin-transfected DC maintained their normal CCR7-mediated migration capacity, and their ability to prime and expand functional cytotoxic T cells recognizing MelanA. Most importantly, E/L-selectin-RNA-transfected DC gained the capability to attach to and roll on sialyl-LewisX in vitro.

Outlook

The presented strategy can be readily translated into the clinic, as it involves no stable genetic manipulation or viral transformation, and allows targeting of a large number of LN.

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Acknowledgments

We thank Dr. Manfred Lutz for fruitful discussions. We thank Drs. Sandy King and Robert Fuhlbrigge for performing pilot rolling experiments. We thank Susanne Rößner and Stefanie Baumann for technical assistance. This study was supported by grants to G.S. from the DFG—German Research Foundation (Collaborative Research Centre SFB643, Project C1), the European Union (DCVACC, contract no.: 503037), and by the Cancer Immunotherapy (CIMT) EU Integrated Project, WP02.02.

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Authors and Affiliations

  1. Department of Dermatology, University Hospital Erlangen, Hartmannstr. 14, 91052, Erlangen, Germany

    Jan Dörrie, Niels Schaft, Ina Müller, Verena Wellner, Tanja Schunder, Jens Hänig, Eckhart Kämpgen & Gerold Schuler

  2. Rudolf Virchow Center, DFG Research Center for Experimental Biomedicine and Department of Dermatology, University of Würzburg, Würzburg, Germany

    Gertie J. Oostingh & Michael P. Schön

  3. Institute Gustave Roussy, Villejuif Cedex, France

    Caroline Robert

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  1. Jan Dörrie
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Correspondence to Niels Schaft.

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Jan Dörrie and Niels Schaft contributed equally to this work.

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Dörrie, J., Schaft, N., Müller, I. et al. Introduction of functional chimeric E/L-selectin by RNA electroporation to target dendritic cells from blood to lymph nodes. Cancer Immunol Immunother 57, 467–477 (2008). https://doi.org/10.1007/s00262-007-0385-1

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