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Lentiviral Vector-Mediated Genetic Programming of Mouse and Human Dendritic Cells

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Genetic Modification of Hematopoietic Stem Cells

Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 506))

Summary

Dendritic cells (DCs) play a key role in the orchestration of immune reactions. Manipulation of DC function through genetic manipulation for vaccine development provides a multitude of applications for active immunotherapy of cancer and chronic infections. Several laboratories have shown that lentiviral vectors (LVs) are efficient and consistent tools for ex vivo gene manipulation of DCs and their precursors. LVs integrate in the genome of target cells resulting in persistent and stable transgene expression, and gene delivery does not result in cytostatic or nonspecific adverse immunomodulatory reactions. Mouse, macaque, and human DCs are efficiently transduced with LVs, allowing preclinical vaccination studies to be gradually implemented into clinical trials. This chapter describes HIV-1-derived LV transduction used for ex vivo gene delivery of marking genes, antigens, and immunomodulatory molecules into mouse and human hematopoietic precursors and DCs. With the perspective of bioengineering DCs from the inside-out, we also describe a one-hit LV transduction method for constitutive expression of GM-CSF and IL-4 genes, which allows self-differentiation of mouse and human hematopoietic precursor cells into highly viable and potent DCs.

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Acknowledgments

The author would like to thank the previous and current members of her group and of the UCLA Vector Core for the hard work and support. This work was supported by The Margareth E. Early Research Trust, Stop Cancer, and NIH grants (UCLA Center for in vivo Imaging in Cancer Biology/2P50-CA086306-06, Rebirth/DFG and SFB 738/DFG.

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Authors and Affiliations

  1. Department of Hematology, Oncology, Hemostaseology, and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany

    Renata Stripecke

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  1. Renata Stripecke
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Editors and Affiliations

  1. Department of Experimental Hematology, Hannover Medical School, Carl-Neuberg-Straβe 1, Hannover, 30625, Germany

    Christopher Baum

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Stripecke, R. (2009). Lentiviral Vector-Mediated Genetic Programming of Mouse and Human Dendritic Cells. In: Baum, C. (eds) Genetic Modification of Hematopoietic Stem Cells. Methods In Molecular Biology™, vol 506. Humana Press. https://doi.org/10.1007/978-1-59745-409-4_11

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  • DOI: https://doi.org/10.1007/978-1-59745-409-4_11

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-980-2

  • Online ISBN: 978-1-59745-409-4

  • eBook Packages: Springer Protocols

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