Abstract
The study of immunometabolism is an important and emerging field in immunology. B-cell activation upon antigen recognition induces profound metabolic changes in the cell, leading to an increase in ATP production to sustain cell proliferation and differentiation. Current methods available to determine the amount of ATP are time-consuming, require extensive sample processing, and need a large amount of starting material. We set up an easy follow-up protocol to determine the relative amount of ATP in living cells, combining cell surface staining with quinacrine. This acridine dye emits a green fluorescent signal in the presence of intracellular ATP. This protocol allows us to determine ATP in small populations of cells using flow cytometry, such as the germinal center.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cyster JG, Allen CDC (2019) B cell responses: cell interaction dynamics and decisions. Cell 177:524–540
Petersone L, Edner NM, Ovcinnikovs V, Heuts F, Ross EM, Ntavli E et al (2018) T cell/B cell collaboration and autoimmunity: an intimate relationship. Front Immunol 9:1941
Mossadegh-Keller N, Brisou G, Beyou A, Nadel B, Roulland S (2021) Human B lymphomas reveal their secrets through genetic mouse models. Front Immunol 12:683597
Jellusova J (2020) Metabolic control of B cell immune responses. Curr Opin Immunol 63:21–28
Cosenza M, Civallero M, Marcheselli L, Sacchi S, Pozzi S (2020) Citarinostat and momelotinib co-target HDAC6 and JAK2/STAT3 in lymphoid malignant cell lines: a potential new therapeutic combination. Apoptosis 25:370–387
Jitschin R, Hofmann AD, Bruns H, Giessl A, Bricks J, Berger J, Saul D, Eckart MJ, Mackensen A, Mougiakakos D (2014) Mitochondrial metabolism contributes to oxidative stress and reveals therapeutic targets in chronic lymphocytic leukemia. Blood 123:2663–2272
Weisel FJ, Mullett SJ, Elsner RA, Menk AV, Trivedi N et al (2020) Germinal center B cells selectively oxidize fatty acids for energy while conducting minimal glycolysis. Nat Immunol 21:331–342
Vessey KA, Ho T, Jobling AI, Wang AY, Fletcher EL (2020) Fluorescent labeling and quantification of vesicular ATP release using live cell imaging. In: Pelegrín P (ed) Purinergic signaling. Methods in molecular biology, vol 2041. Humana, New York
Peti-Peterdi J, Fintha A, Fuson AL, Tousson A, Chow RH (2004) Real-time imaging of renin release in vitro. Am J Physiol Renal Physiol 287:F329–F335
Acknowledgments
This manuscript was funded by grant RTI2018-101586-A-100 from the Spanish Ministry of Science, Innovation and Universities.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Iborra Pernichi, M., Ruiz García, J., Martínez-Martín, N. (2023). Quantification of Intracellular ATP Content in Ex Vivo GC B Cells. In: Papa, S., Bubici, C. (eds) Metabolic Reprogramming. Methods in Molecular Biology, vol 2675. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3247-5_9
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3247-5_9
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3246-8
Online ISBN: 978-1-0716-3247-5
eBook Packages: Springer Protocols