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| Status |
Public on Jul 01, 2012 |
| Title |
Transcriptional profiling analysis of pre-selected DP thymocytes in response to positive and negative selection signals |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
|
| Summary |
Sustained Ca2+ entry into CD4+CD8+ double-positive thymocytes is required for positive selection. We identified a voltage-gated Na+ channel (VGSC), essential for positive selection of CD4+ T cells. Pharmacological inhibition of VGSC activity inhibited sustained Ca2+ influx induced by positive-selecting ligands and in vitro positive selection of CD4+ but not CD8+ T cells. In vivo shRNA knockdown of Scn5a specifically inhibited positive selection of CD4+ T cells. Ectopic expression of VGSC in peripheral AND CD4+ T cells bestowed the ability to respond to a positively selecting ligand, directly demonstrating VGSC expression was responsible for increased sensitivity. Thus active VGSCs in thymocytes provides a mechanism by which a weak positive selecting signal can induce sustained Ca2+ signals required for CD4+ T cell development.
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| Overall design |
Pre-selected AND DP thymocytes were stimulated with plate bound peptide-loaded I-Ek Ig dimers. Genes were differentially regulated by positive and negative selection signals.
The goal of this study is to identify ion-channel related genes critical for thymic positive selection, given the sustained Ca2+ entry into double positive thymocytes is required for positive selection. Total RNA of pre-selected AND thymocytes stimulated with I-Ek Ig dimers loaded with the positively-selecting peptide gp250, the negatively-selecting agonist peptide MCC, or the non-selecting control peptide Hb were used for the transcriptional profiling analysis. We found 28 genes were differentially down-regulated by MCC stimulation but upregulated by gp250 stimulation. One of them, scn4b, was ion-channel related.
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| Contributor(s) |
Lo W, Allen PM |
| Citation(s) |
22842345 |
| Submission date |
Jun 25, 2012 |
| Last update date |
Jun 14, 2018 |
| Contact name |
Wan-Lin Lo |
| Organization name |
Washington University in St Louis
|
| Street address |
425 S Euclid Ave, campus box 8118
|
| City |
Saint Louis |
| ZIP/Postal code |
63110 |
| Country |
USA |
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| Platforms (1) |
| GPL6885 |
Illumina MouseRef-8 v2.0 expression beadchip |
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| Samples (6)
|
| GSM952008 |
AND.Rag1-/-H-2d DP thymocytes_MCC/I-Ek stimulation_rep1 |
| GSM952009 |
AND.Rag1-/-H-2d DP thymocytes_gp250/I-Ek stimulation_rep1 |
| GSM952010 |
AND.Rag1-/-H-2d DP thymocytes_Hb/I-Ek stimulation_rep1 |
| GSM952011 |
AND.Rag1-/-H-2d DP thymocytes_MCC/I-Ek stimulation_rep2 |
| GSM952012 |
AND.Rag1-/-H-2d DP thymocytes_gp250/I-Ek stimulation_rep2 |
| GSM952013 |
AND.Rag1-/-H-2d DP thymocytes_Hb/I-Ek stimulation_rep2 |
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| Relations |
| BioProject |
PRJNA169320 |
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