|
| Status |
Public on Mar 01, 2009 |
| Title |
Activated T cells 4 |
| Sample type |
RNA |
| |
|
| Source name |
Peripheral CD4+Foxp3-EGFP- T cells treated with anti-CD3+anti-CD28 mAbs
|
| Organism |
Mus musculus |
| Characteristics |
Foxp3/EGFP bicistronic mice on BALB/c background
|
| Treatment protocol |
Group 1: induced regulatory T cells, derived by treatment of CD4+Foxp3-EGFP– T cells of Foxp3EGFP bicistronic mice with anti-CD3+anti-CD28 monoclonal antibodies (mAbs) +TGF-beta for 3 days, then sorting for CD4+Foxp3+EGFP+ cells; Group 2: Failed induced regulatory T cells: CD4+Foxp3-EGFP– cells, derived from group 1 cultures that failed to express Foxp3EGFP following treatment with anti-CD3+anti-CD28 mAbs+TGF-beta; Group 3: induced CD4+Foxp3deltaEGFP+ cells, derived by treatment of CD4+Foxp3deltaEGFP– T cells of Foxp3deltaEGFP mutant mice with anti-CD3+anti-CD28 mAbs+TGF-beta for 3 days, then sorting for CD4+Foxp3deltaEGFP+ cells; Group 4: activated T cells, derived from conventional CD4+Foxp3-EGFP- T cells of Foxp3/EGFP bicistronic mice that were stimulated in vitro with anti-CD3+anti-CD28 mAbs but NO TGF-beta; Group 5: Activated natural regulatory T cells are CD4+Foxp3+EGFP+ T cells isolated from Foxp3/EGFP bicistronic mice and activated with anti-CD3+antiCD28 mAbs +TGF-beta to simulate treatment of groups 1-3; Group 6: native (not activated) CD4+Foxp3+EGFP+ natural regulatory T cells of Foxp3EGFP bicistronic mice; Group 7: Native (not activated) CD4+Foxp3deltaEGFP+ T cells isolated from heterozygos Foxp3deltaEGFP females; Group 8: Native (not activated) conventional CD4+T cells isolated from bicistonic Foxp3/EGFP mice; Group 9: IL-2-cultured induced regulatory T cells, derived by treatment of CD4+Foxp3-EGFP– T cells (from Foxp3EGFP bicistronic muce) with anti-CD3+anti-CD28 mAbs +TGF-beta for 3 days then sorting for CD4+Foxp3+EGFP+ cells and culturing those for one week with IL-2 at 100 units/ml; Group 10: IL-2-cultured induced CD4+Foxp3deltaEGFP+ cells, derived by treatment of CD4+Foxp3deltaEGFP– T cells (isolated from Foxp3deltaEGFP mice) with anti-CD3+anti-CD28 mAbs+TGF-beta for 3 days, then sorting for CD4+Foxp3deltaEGFP+ cells and culturing those for one week with IL-2 at 100 units/ml.
|
| Growth protocol |
Conventional CD4+Foxp3-EGFP- and natural CD4+Foxp3+EGFP+ regulatory T cells were isolated from secondary lymphoid tissues (spleens and lymph nodes) of bicistronic Foxp3/EGFP mice. Conventional CD4+Foxp3deltaEGFP- were isolated from Foxp3deltaEGFP mutant males, while CD4+Foxp3deltaEGFP+ regulatory T cell precursors were isolated from heterozygous Foxp3deltaEGFP females. The respective cell populations were isolated by cell sorting.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated by Trizol extraction and was furher purified by Qiagen Rneasy spin columns following the manufacturer's specifications
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol
|
| |
|
| Hybridization protocol |
Following fragmentation, cRNA were hybridized for 16 hr at 45C on Affymetrix GeneChip Mouse Genome 430 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Group 4
|
| Data processing |
The data was normalized using the justRMA algorithm from the Bioconductor group following the methods of the Celsius project.
|
| |
|
| Submission date |
Jan 14, 2009 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Brian O'Connor |
| Organization name |
UCLA
|
| Street address |
2018 Griffith Park Blvd
|
| City |
los angeles |
| State/province |
CA |
| ZIP/Postal code |
90039 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE14415 |
Gene expression profiling of natural and induced regulatory T cells |
|
| Relations |
| Reanalyzed by |
GSE119085 |
