Abstract
Affinity maturation of B cell clones within germinal centers constitutes an important mechanism for immune memory. During this process, B cell receptor signaling capacity is tested in multiple rounds of positive selection. Antigen stimulation and co-stimulatory signals mobilize calcium to switch on gene expression leading to proliferation and survival and to differentiation into memory B cells and plasma cells. Additionally, all these processes require adaption of B cell metabolism, and calcium signaling and metabolic pathways are closely interlinked. Mitochondrial adaption, ROS production, and NADPH oxidase activation are involved in cell fate decisions, but it remains elusive to what extent, especially because the analysis of these dynamic processes in germinal centers has to take place in vivo. Here, we introduce a quantitative intravital imaging method for combined measurement of cytoplasmic calcium concentration and enzymatic fingerprinting in germinal center B cells as a possible tool in order to further examine the relationship of calcium signaling and immunometabolism.
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Acknowledgments
We thank Robert Günther, Ralf Uecker, and Peggy Mex for excellent technical assistance. A.E.H. was supported by Deutsche Forschungsgemeinschaft (DFG) collaborative research grant TRR130, project P17, and by grant HA5354/12-1. A.E.H. and R.A.N. were supported DFG TRR130, project C01 and DFG CRC1444, project 14, and a grant from the Einstein Foundation Berlin (A-2019-559).
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Ulbricht, C., Leben, R., Cao, Y., Niesner, R.A., Hauser, A.E. (2023). Combined FRET-FLIM and NAD(P)H FLIM to Analyze B Cell Receptor Signaling Induced Metabolic Activity of Germinal Center B Cells In Vivo. In: Baldari, C.T., Dustin, M.L. (eds) The Immune Synapse. Methods in Molecular Biology, vol 2654. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3135-5_6
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