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In Vivo Modulation of Gene Expression by Lentiviral Transduction in “Human Immune System” Rag2−/−γc −/− Mice

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Dendritic Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 595))

Abstract

Over the last two decades, several humanized mouse models have been used to experimentally analyze the function and development of the human immune system. Recent advances have lead to the establishment of new murine−human chimeric models with improved characteristics, both in terms of human engraftment efficiency and in situ multilineage human hematopoietic development. We describe here the use of newborn BALB/c Rag2−/−γc −/− mice as recipients of human hematopoietic progenitor cells to produce “human immune system” (HIS) (BALB-Rag/γ) mice, using human fetal liver progenitors. The two major subsets of the human dendritic cell lineage, namely, BDCA2+CD11c− plasmacytoid dendritic cells and BDCA2−CD11c+ conventional dendritic cells, can be found in HIS (BALB-Rag/γ) mice. In order to manipulate the expression of genes of interest, the human hematopoietic progenitor cells can be genetically engineered ex vivo by lentiviral transduction before performing xenograft transplantation. Using this mouse model, the human immune system can be assessed for both fundamental and pre-clinical purposes.

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Acknowledgments

We thank the staff of the ABSL-3 unit of the Animal Research Institute Amsterdam (ARIA) located at the Academic Medical Center (AMC) of Amsterdam for excellent care to the animals. We also thank Berend Hooibrink for expertise in cell sorting and maintenance of the flow cytometry facility. Last, we are grateful to the Bloemenhove Clinic (Heemstede, The Netherlands) and the Department of Obstetrics and Gynecology of the AMC for providing fetal tissues and umbilical cord blood, respectively. Mireille Centlivre is supported by a Marie-Curie Intra-European fellowship (MEIF-C-2007-039689). This work is supported by the Grand Challenges in Global Health Initiative (Bill & Melinda Gates Foundation, Wellcome Trust, Foundation for the National Institute of Health, Canadian Institutes of Health Research) as part of the “Human Vaccine Consortium”, the Dutch society for scientific research (NWO), the Dutch Foundation for Cancer Research (KWF), and the Landsteiner Blood Transfusion Research Foundation (LSBR).

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Authors and Affiliations

  1. Department of Cell Biology and Histology, Center for Immunology Amsterdam (CIA), Academic Medical Center of the University of Amsterdam (AMC-UvA), Amsterdam, The Netherlands

    Anja U. van Lent, Maho Nagasawa, Julien J. Karrich, Stephan M. Pouw, Kees Weijer, Hergen Spits, Bianca Blom & Nicolas Legrand

  2. Laboratory of Experimental Virology, Department of Cell Biology and Histology, Center for Immunology Amsterdam (CIA), Center for Infection and Immunity Amsterdam (CINIMA), Amsterdam, The Netherlands

    Mireille Centlivre

  3. Academic Medical Center of the University of Amsterdam (AMC-UvA), Amsterdam, The Netherlands

    Mireille Centlivre

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  1. Anja U. van Lent
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  2. Mireille Centlivre
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  3. Maho Nagasawa
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  4. Julien J. Karrich
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  5. Stephan M. Pouw
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  6. Kees Weijer
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  7. Hergen Spits
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  8. Bianca Blom
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  9. Nicolas Legrand
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  1. Quellijnstraat 25A, Amsterdam, 1072 XN, Netherlands

    Shalin H. Naik

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van Lent, A.U. et al. (2010). In Vivo Modulation of Gene Expression by Lentiviral Transduction in “Human Immune System” Rag2−/−γc −/− Mice. In: Naik, S. (eds) Dendritic Cell Protocols. Methods in Molecular Biology, vol 595. Humana Press. https://doi.org/10.1007/978-1-60761-421-0_6

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  • DOI: https://doi.org/10.1007/978-1-60761-421-0_6

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