Abstract
Deletion of autoreactive thymocytes at the DP stage is the basis for tolerance to thymus-expressed self antigens. In this study we investigated whether distinct signalling pathways are induced in DP thymocytes as compared to mature T cells upon stimulation with antigen. Using triple transgenic mice expressing a TCR transgene, dominant negative ras/Mek proteins and a reporter gene construct with AP-1 or NF-κB binding sites, we showed a complete lack of transcriptional activity of NF-κB but not AP-1 in DP thymocytes, whereas both were transcriptionally active in mature T cells after antigenic stimulation. Lack of NF-κB induction correlated with increased death in response to antigen. AP-1 induction was dependent on the integrity of the ras/Mek pathway indicating that this pathway was activated in the DP thymocytes. In contrast, we found a complete lack of constitutive expression of the ε isoform of Protein Kinase C (PKC) in DP thymocytes, although it was present in mature thymocytes and peripheral T cells. Taken together the results suggest that the lack of PKCε in DP thymocytes could lead to the absence of NF-κB activity after antigenic stimulation contributing to negative selection.
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Abbreviations
- PKC:
-
protein kinase C
- TCR:
-
T cell receptor
- DP:
-
double positive thymocytes
- SP:
-
single positive thymocytes
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Acknowledgements
We thank José Alberola-Ila and Roger Perlmutter for providing the dnras/dMek mice, Peter Parker for the PKC antisera, Gilles Warcollier and Michel Pontier for animal care and Sylvie Guerder for providing data on peripheral T cells, performing Oct-1 gelshifts and breeding the luciferase mice. AK Simon was a recipient of a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft. This work was supported by institutional grants from Institut National de la Santé et de la Recherche médicale (INSERM) and Centre National de la Recherche Scientifique (CNRS) and by grants from Association pour la Recherche sur le Cancer (ARC), the Ligue Nationale Contre le Cancer (LNCC) and the LNCC-Comité des Bouches du Rhône.
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Simon, A., Auphan, N., Pophillat, M. et al. The lack of NF-κB transactivation and PKCε expression in CD4+CD8+ thymocytes correlates with negative selection. Cell Death Differ 7, 1253–1262 (2000). https://doi.org/10.1038/sj.cdd.4400760
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DOI: https://doi.org/10.1038/sj.cdd.4400760
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