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| Status |
Public on Dec 14, 2011 |
| Title |
Dominant Th2 differentiation of human regulatory T cells upon loss of FOXP3 expression |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
CD4+CD25+FOXP3+ regulatory T cells (Treg) are pivotal for peripheral self-tolerance. They prevent immune responses to auto- and alloantigens and are thus under close scrutiny as cellular therapeutics for autoimmune diseases and the prevention or treatment of alloresponses after organ or stem cell transplantation. We previously showed that human Treg cells with a memory cell phenotype, but not those with a naïve phenotype, rapidly down-regulate expression of the lineage-defining transcription factor forkhead box P3 (FOXP3) upon in vitro expansion. We now compared the transcriptomes of stable FOXP3+ Treg and converted FOXP3- 'ex-Treg' cells by applying a newly developed intranuclear staining protocol that permits the isolation of intact mRNA from fixed, permeabilized and FACS-purified cell populations. Whole genome microarray analysis revealed strong and selective upregulation of Th2 signature genes, including GATA-3, IL-4, IL-5 and IL-13, upon downregulation of FOXP3. Th2 differentiation of converted, FOXP3- ex-Treg cells occurred even under non-polarizing conditions and could not be prevented by IL-4 signaling blockade. Thus, our studies identify Th2 differentiation as the default developmental program of human Treg cells after downregulation of FOXP3.
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| Overall design |
RNA preparations from FOXP3 stained in vitro expanded CD4+CD25highCD45RA- “memory” Treg cells from five independent donors were analyzed using Whole Human Genome Oligo Microarrays (Agilent). An adapted FOXP3 staining procedure to stain FOXP3 in human regulatory T cells to isolate intact RNA for microarray hybridization was developed (see extract protocol).
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| Contributor(s) |
Hansmann L, Schmidl C, Andreesen R, Kett J, Steger L, Hoffmann P, Rehli M, Edinger M |
| Citation(s) |
22210907 |
| Submission date |
Dec 20, 2010 |
| Last update date |
Nov 27, 2018 |
| Contact name |
Christian Schmidl |
| E-mail(s) |
christian.schmidl@klinik.uni-regensburg.de
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| Organization name |
Rengensburg Center for Interventional Immunology
|
| Lab |
Schmidl Lab
|
| Street address |
Franz-Josef-Strauss-Allee 11
|
| City |
Regensburg |
| ZIP/Postal code |
93053 |
| Country |
Germany |
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| Platforms (1) |
| GPL13607 |
Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Feature Number version) |
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| Samples (10)
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| Relations |
| BioProject |
PRJNA135017 |
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