Lck Mediates Signal Transmission from CD59 to the TCR/CD3 Pathway in Jurkat T Cells
Anna
M.
Lipp1,
Kata
Juhasz1,
Christian
Paar1, 2,
Christoph
Ogris3,
Paul
Eckerstorfer4,
Roland
Thuenauer1, 5,
Jan
Hesse1,
Hannes
Stockinger4,
Gerhard
Schütz6,
Ulrich
Bodenhofer3,
Zsolt
Balogi1* and
Alois
Sonnleitner1*
-
1
Center for Advanced Bioanalysis GmbH, Austria
-
2
General Hospital Linz, Austria
-
3
Johannes Kepler University Linz, Institute for Bioinformatics, Austria
-
4
Medical University Vienna, Molecular Immunology Unit, Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Austria
-
5
Albert-Ludwigs- Universität Freiburg, BIOSS Centre for Biological Signalling Studies, Germany
-
6
Vienna University of Technology, Institute of Applied Physics, Austria
Functional heterogeneity has been considered as a principle mechanism of the adaptive immune response. Here we developed an imaging-based method for studying Ca2+ signaling at the single-cell level as a function of time and stimulus. Jurkat T cells were exposed to stimulatory anti-CD3ε- or anti-CD59-coated surfaces and individual Ca2+ time traces were analyzed via unsupervised clustering. This analysis revealed a heterogeneous Ca2+ response of the cell population in a stimulus-dependent manner. Interestingly, an unresponsive subpopulation identified upon anti-CD59 stimulation coincided with a low CD3 surface expressing subpopulation observed in flow cytometry measurements. Further analysis of TCR/CD3 deficient or overexpressing cells showed that CD59-mediated signaling is strongly dependent on TCR/CD3 surface expression. However, in protein co-patterning and fluorescence recovery after photobleaching experiments no direct physical interaction was observed between CD59 and CD3 at the plasma membrane upon anti-CD59 stimulation. Nonetheless, siRNA-mediated protein knock-downs of downstream signaling molecules revealed that the Src family kinase Lck is essential for both signaling pathways. Moreover, physically linking Lck to CD3ζ completely abolished CD59-triggered Ca2+ signaling, while signaling was still functional upon direct TCR/CD3 stimulation. Altogether, we demonstrate that Lck mediates signal transmission from CD59 to the TCR/CD3 pathway in Jurkat T cells, and propose that CD59 may act via Lck to modulate T cell responses.
Keywords:
CD3,
CD59,
Lck,
Calcium Signaling,
TCR signaling
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Immune receptors and signaling
Citation:
Lipp
AM,
Juhasz
K,
Paar
C,
Ogris
C,
Eckerstorfer
P,
Thuenauer
R,
Hesse
J,
Stockinger
H,
Schütz
G,
Bodenhofer
U,
Balogi
Z and
Sonnleitner
A
(2013). Lck Mediates Signal Transmission from CD59 to the TCR/CD3 Pathway in Jurkat T Cells.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.01145
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Received:
17 Jul 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Dr. Zsolt Balogi, Center for Advanced Bioanalysis GmbH, Linz, Austria, zsolt.balogi@cbl.at
Dr. Alois Sonnleitner, Center for Advanced Bioanalysis GmbH, Linz, Austria, alois.sonnleitner@cbl.at