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| Status |
Public on Sep 30, 2016 |
| Title |
Celiac disease T cell clone response to CD3/CD28 stimulation |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
To identify the CD4+ T cell cytokines responsible for the proliferation of the Lin-IEL lines CD4+ T cell clone L10, which recognises DQ2-glia-α1, one of the immunodominant T cell epitopes in celiac disease, was stimulated for 3 hours in IMDM with plate-bound CD3/CD28-specific (2.5 µg/ml each) or control antibodies coated onto 6-well non-tissue culture treated plates. Three independent biological replicates were performed, each time including 6 million Ficoll-purified live cells per condition. RNA was purified from these cells using the RNAeasy mini kit (Qiagen, Venlo, the Netherlands). cDNA was amplified using the Applause WT-Amp system (NuGEN technologies, Bemmel, the Netherlands) and biotin-labelled with the Encore Biotin Module (NuGEN). Human Gene 1.0 ST arrays (Affymetrix, High Wycombe, UK) were employed to quantify global gene expression.
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| Overall design |
CD4+ T clone L10 cells, isolated from a duodenal biopsy of a celiac disease patient (Petersen et al., 2014, Nat. Struct. Biol. 21:480-488) were stimulated for 3 hours in IMDM with plate-bound CD3/CD28-specific (2.5 µg/ml each) or control antibodies. Three independent biological replicates were performed, each time including 6 million Ficoll-purified live cells per condition.
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| Contributor(s) |
Brugman MH, van Bergen J |
| Citation(s) |
28049849 |
| Submission date |
Sep 30, 2015 |
| Last update date |
Jul 26, 2018 |
| Contact name |
Martijn H Brugman |
| E-mail(s) |
m.h.brugman@lumc.nl
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| Phone |
+31-71-5261300
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| Organization name |
LUMC
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| Department |
IHB
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| Street address |
Albinusdreef 2
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| City |
Leiden |
| ZIP/Postal code |
2333 ZA |
| Country |
Netherlands |
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| Platforms (1) |
| GPL6244 |
[HuGene-1_0-st] Affymetrix Human Gene 1.0 ST Array [transcript (gene) version] |
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| Samples (6)
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| GSM1898773 |
CD4+Tcell_clone_L10_unstimulated_replicate1 |
| GSM1898774 |
CD4+Tcell_clone_L10_CD3CD28_replicate1 |
| GSM1898775 |
CD4+Tcell_clone_L10_unstimulated_replicate2 |
| GSM1898776 |
CD4+Tcell_clone_L10_CD3CD28_replicate2 |
| GSM1898777 |
CD4+Tcell_clone_L10_unstimulated_replicate3 |
| GSM1898778 |
CD4+Tcell_clone_L10_CD3CD28_replicate3 |
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| Relations |
| BioProject |
PRJNA297406 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE73599_RAW.tar |
25.0 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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