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Simultaneous In Vivo Monitoring of Regulatory and Effector T Lymphocytes Using Secreted Gaussia Luciferase, Firefly Luciferase, and Secreted Alkaline Phosphatase

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Bioluminescent Imaging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1098))

Abstract

Regulatory T cells (Tregs) are amongst the most widely studied cells in a variety of immune-mediated conditions, including transplantation and Graft Versus Host Disease (GVHD), cancer and autoimmunity; indeed, there is great interest in the tolerogenic potential of Treg-based therapy. Consequently, the need to establish the mechanisms that determine Treg survival and longevity, in addition to developing new tools to monitor these parameters, is paramount. Using both a mouse model of GVHD and a mouse model of Type 1 Diabetes (T1D), we describe herein a dual reporter system based on Gluc and multiplexed with SEAP and non-secreted Firefly luciferase (Fluc), which permits simultaneous imaging and noninvasive tracking of two different T-cell populations (CD4+CD25+ Tregs and CD4+CD25− Tcon cells) in vivo by transducing the cells with different lentiviruses bearing distinct color signatures. This new technology promises to overcome the limitations of the conventional methods currently available to study lymphocyte survival in vivo. Furthermore, this novel technique has applications not only in autoimmunity and alloimmunity, but also in the wider field of immunology.

Grant K. Lewandrowski and Ciara N. Magee contributed equally to this work.

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Author information

Authors and Affiliations

  1. Experimental Therapeutics and Molecular Imaging Laboratory, Department of Neurology, Neuroscience Center, Massachusetts General Hospital, Boston, MA, USA

    Grant K. Lewandrowski & Bakhos A. Tannous

  2. Program in Neuroscience, Harvard Medical School, Boston, MA, USA

    Grant K. Lewandrowski & Bakhos A. Tannous

  3. Renal Division, Transplantation Research Center, Brigham and Women’s Hospital and Children’s Hospital, Harvard Medical School, Boston, MA, USA

    Ciara N. Magee, Marwan Mounayar & Jamil Azzi

Authors
  1. Grant K. Lewandrowski
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  2. Ciara N. Magee
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  3. Marwan Mounayar
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  4. Bakhos A. Tannous
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  5. Jamil Azzi
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Editor information

Editors and Affiliations

  1. Department of Neurology, Neuroscience Center, Massachusetts General Hospital, Boston, Massachusetts, USA

    Christian E. Badr

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Lewandrowski, G.K., Magee, C.N., Mounayar, M., Tannous, B.A., Azzi, J. (2014). Simultaneous In Vivo Monitoring of Regulatory and Effector T Lymphocytes Using Secreted Gaussia Luciferase, Firefly Luciferase, and Secreted Alkaline Phosphatase. In: Badr, C. (eds) Bioluminescent Imaging. Methods in Molecular Biology, vol 1098. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-718-1_17

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  • DOI: https://doi.org/10.1007/978-1-62703-718-1_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-717-4

  • Online ISBN: 978-1-62703-718-1

  • eBook Packages: Springer Protocols

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