Lck mediates signal transmission from CD59 to the TCR/CD3 pathway in Jurkat T cells
| Autor: | Alois Sonnleitner, Paul Eckerstorfer, Benedikt Nimmervoll, Gerhard J. Schütz, Zsolt Balogi, Kata Juhasz, Roland Thuenauer, Anna M. Lipp, Ulrich Bodenhofer, Hannes Stockinger, Christoph Ogris, Jan Hesse, Christian Paar |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
CD3 Complex
CD3 T cell Immune Cells Immunology Receptors Antigen T-Cell lcsh:Medicine chemical and pharmacologic phenomena CD59 Antigens Signal transduction Jurkat cells Signaling Pathways Jurkat Cells Molecular cell biology medicine Calcium-Mediated Signal Transduction Humans Signaling in Cellular Processes Membrane Receptor Signaling Src family kinase Calcium Signaling lcsh:Science Biology Multidisciplinary biology TCR signaling cascade T Cells ZAP70 T-cell receptor lcsh:R Cell Membrane Mechanisms of Signal Transduction CD28 Signaling cascades hemic and immune systems Cell biology medicine.anatomical_structure Lymphocyte Specific Protein Tyrosine Kinase p56(lck) biology.protein Medicine lcsh:Q Clinical Immunology Cellular Types Immunologic Receptor Signaling Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 9, Iss 1, p e85934 (2014) |
| ISSN: | 1932-6203 |
| Popis: | The glycosylphosphatidylinositol (GPI)-anchored molecule CD59 has been implicated in the modulation of T cell responses, but the underlying molecular mechanism of CD59 influencing T cell signaling remained unclear. Here we analyzed Jurkat T cells stimulated via anti-CD3ε- or anti-CD59-coated surfaces, using time-resolved single-cell Ca(2+) imaging as a read-out for stimulation. This analysis revealed a heterogeneous Ca(2+) response of the cell population in a stimulus-dependent manner. Further analysis of T cell receptor (TCR)/CD3 deficient or overexpressing cells showed that CD59-mediated signaling is strongly dependent on TCR/CD3 surface expression. 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. However, siRNA-mediated protein knock-downs of downstream signaling molecules revealed that the Src family kinase Lck and the adaptor molecule linker of activated T cells (LAT) are essential for both signaling pathways. Furthermore, flow cytometry measurements showed that knock-down of Lck accelerates CD3 re-expression at the cell surface after anti-CD59 stimulation similar to what has been observed upon direct TCR/CD3 stimulation. Finally, physically linking Lck to CD3ζ completely abolished CD59-triggered Ca(2+) 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. |
| Databáze: | OpenAIRE |
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