PMID: 11933010Apr 5, 2002Paper

Flow cytometry used for the analysis of calcium signaling induced by antigen-specific T-cell activation

Cytometry
Bence RéthiE Rajnavölgyi

Abstract

In this study, the effect of antigen-presenting cells (APC), peptide concentration, and CD28 costimulation on calcium signaling, induced by antigen-specific T-cell activation, was studied by flow cytometry. We used two experimental approaches, which differed in their time scale and in the duration of the T cell-APC interaction, to measure the increase of intracellular free calcium levels ([Ca(2+)](i)) in activated T cells: (1) Fluo-3-loaded T cells were activated by cocentrifugation with peptide-loaded APC and the kinetics of fluorescence intensity changes was monitored continuously and (2) peptide-loaded APC and T cells were mixed, cocultured, and the fluorescence intensity was measured at various time intervals. The calcium signal of T cells was dependent on the APC as demonstrated by the ratio of cells exhibiting high versus low fluorescence intensity and by the magnitude of the calcium signal in the activated population. Short-term interaction of T cells with less potent APC or with efficient APC in the presence of low antigen concentration resulted in decreased calcium signaling. CD28-mediated costimulation enhanced the magnitude and sustained the increase of intracellular calcium levels. In line with the strong and sustai...Continue Reading

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Citations

Aug 4, 2011·Proceedings of the National Academy of Sciences of the United States of America·Lina ZhangKexin Chen
Mar 27, 2007·Annals of the New York Academy of Sciences·Endre KissJános Matkó
Apr 1, 2015·Immunology and Cell Biology·Susan N ChristoJohn D Hayball
Jan 1, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·A S KaposiA Treszl
Jul 29, 2005·Toxicological Sciences : an Official Journal of the Society of Toxicology·Jun GaoScott W Burchiel
Feb 8, 2003·The Journal of Immunology : Official Journal of the American Association of Immunologists·Harnisha DalwadiJonathan Braun
Jan 29, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Michael E March, Eric O Long
Jan 1, 2014·The Journal of Immunology : Official Journal of the American Association of Immunologists·Andrés TittarelliFlavio Salazar-Onfray
Feb 9, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Mythili KoneruAlan B Frey
May 26, 2004·Cellular Signalling·Katalin LudanyiEva Rajnavolgyi

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