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Radioisotope-Based Protocol for Determination of Central Carbon Metabolism in T Cells

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T-Cell Receptor Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2111))

Abstract

T lymphocytes are the major components of the adaptive immune system. It’s been known that T cells are able to engage a diverse range of metabolic programs to meet the metabolic demands during their life cycle from early development, activation to functional differentiation. Central carbon metabolic pathways provide energy, reducing power, and biosynthetic precursors to support T cell homeostasis, proliferation, and immune functions. As such, quantitative or semiquantitative analysis of central carbon metabolic flux activities offers mechanistic details, as well as insights into the regulation of metabolic pathways and the impact of changing metabolic programs on T cell life cycle. Global profiling of cellular metabolites by mass spectrometry-based metabolomics and metabolic flux analysis (MFA) using radioactive and nonradioactive/stable isotope approaches are powerful tools for determination of central carbon metabolic pathway activity. Here, we describe in detail the procedure for the radioisotope-based approach of analyzing central carbon metabolic fluxes in T cells.

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Acknowledgments

This work was supported by 1R01AI114581 from the National Institute of Health and 128436-RSG-15-180-01-LIB from the American Cancer Society (R.W.).

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Authors and Affiliations

  1. Center for Childhood Cancer & Blood Diseases, Hematology/Oncology & BMT, The Research Institute at Nationwide Children’s Hospital, Ohio State University, Columbus, OH, USA

    Xuyong Chen, John William Sherman & Ruoning Wang

Authors
  1. Xuyong Chen
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  2. John William Sherman
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  3. Ruoning Wang
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Corresponding author

Correspondence to Ruoning Wang .

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Editors and Affiliations

  1. Department of Pathogen Biology, Huazhong University of Science and Technology, Wuhan, China

    Chaohong Liu

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Chen, X., Sherman, J.W., Wang, R. (2020). Radioisotope-Based Protocol for Determination of Central Carbon Metabolism in T Cells. In: Liu, C. (eds) T-Cell Receptor Signaling. Methods in Molecular Biology, vol 2111. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0266-9_20

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  • DOI: https://doi.org/10.1007/978-1-0716-0266-9_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0265-2

  • Online ISBN: 978-1-0716-0266-9

  • eBook Packages: Springer Protocols

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