Dataset: Retinoic Acid Enhances Foxp3 Induction Indirectly by Relieving Inhibition from CD4(+)CD44(hi) Cells.
CD4(+)Foxp3(+) regulatory T (Treg) cells originate primarily from thymic differentiation, but conversion of mature T lymphocytes to Foxp3...
CD4(+)Foxp3(+) regulatory T (Treg) cells originate primarily from thymic differentiation, but conversion of mature T lymphocytes to Foxp3 positivity can be elicited by several means, including in vitro activation in the presence of TGF-beta. Retinoic acid (RA) increases TGF-beta-induced expression of Foxp3, through unknown molecular mechanisms. We showed here that, rather than enhancing TGF-beta signaling directly in naive CD4(+) T cells, RA negatively regulated an accompanying population of CD4(+) T cells with a CD44(hi) memory and effector phenotype. These memory cells actively inhibited the TGF-beta-induced conversion of naive CD4(+) T cells through the synthesis of a set of cytokines (IL-4, IL-21, IFN-gamma) whose expression was coordinately curtailed by RA. This indirect effect was evident in vivo and required the expression of the RA receptor alpha. Thus, cytokine-producing CD44(hi) cells actively restrain TGF-beta-mediated Foxp3 expression in naive T cells, and this balance can be shifted or fine-tuned by RA. All gene expression profiles were obtained from highly purified T cell populations sorted by flow cytometry. To reduce variability, cells from multiple mice were pooled for sorting, and replicates were generated for essentially all groups. RNA from 0.5-3 x 105 cells was amplified, labeled, and hybridized to Affymetrix M430v2 microarrays. Raw data were preprocessed with the RMA algorithm in GenePattern, and averaged expression values were used for analysis.
- Species:
- mouse
- Samples:
- 17
- Source:
- E-GEOD-13306
- PubMed:
- 19006694
- Updated:
- Dec.12, 2014
- Registered:
- Nov.10, 2014
| Sample | POST-CULTURE SORT | CULTURE | GENETIC BACKGROUND | ORGANISM PART |
|---|---|---|---|---|
| GSM335922 | Sorted on CD4+ Foxp3/GFP- | CD4+Foxp3- T cells were culture with LP-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells were obtained from spleen and dendritic cells were obtained from lamina propria |
| GSM335922 | Sorted on CD4+ Foxp3/GFP- | CD4+Foxp3- T cells were culture with LP-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells were obtained from spleen and dendritic cells were obtained from lamina propria |
| GSM335924 | Sorted on CD4+ Foxp3/GFP+ | CD4+Foxp3- T cells were culture with LP-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells were obtained from spleen and dendritic cells were obtained from lamina propria |
| GSM335924 | Sorted on CD4+ Foxp3/GFP+ | CD4+Foxp3- T cells were culture with LP-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells were obtained from spleen and dendritic cells were obtained from lamina propria |
| GSM335926 | Sorted on CD4+ Foxp3/GFP- | CD4+Foxp3- T cells were culture with Sp-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + RA (100nM) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells and dendritic cells were obtained from spleen |
| GSM335927 | Sorted on CD4+ Foxp3/GFP+ | CD4+Foxp3- T cells were culture with Sp-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + RA (100nM) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells and dendritic cells were obtained from spleen |
| GSM335927 | Sorted on CD4+ Foxp3/GFP+ | CD4+Foxp3- T cells were culture with Sp-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + RA (100nM) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells and dendritic cells were obtained from spleen |
| GSM335929 | Sorted on CD4+ Foxp3/GFP- | CD4+Foxp3- T cells were culture with Sp-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells and dendritic cells were obtained from spleen |
| GSM335929 | Sorted on CD4+ Foxp3/GFP- | CD4+Foxp3- T cells were culture with Sp-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells and dendritic cells were obtained from spleen |
| GSM33593 | Sorted on CD4+ Foxp3/GFP+ | CD4+Foxp3- T cells were culture with Sp-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells and dendritic cells were obtained from spleen |
| GSM33593 | Sorted on CD4+ Foxp3/GFP+ | CD4+Foxp3- T cells were culture with Sp-DCs at a 10:1 ratio with anti-CD3 (1ug/ml) + TGFbeta (0.6ng/ml) + IL-2 (5ng/ml on D2 and D4) for 5 days | C57BL/6-Foxp3/GFP | CD4+Foxp3- T cells and dendritic cells were obtained from spleen |
| GSM335933 | Memory T cells were sorted based on the expression of the congenic marker (CD45.1 or CD45.2) | Congenically marked (CD45.1 or CD45.2) memory T cells were cultured with CD44negCD62LhiCD25-CD4+ (naive) T cells at a 2:1 ratio with anti-CD3/CD28 beads + IL-2 (20 U/ml) + TGFbeta (10 ng/ml) for 48hrs | C57BL/6 | CD44hiCD62L-CD25-CD4+ (Memory) T cells were sorted form spleen and lymph nodes (mesenteric and cervical) |
| GSM335933 | Memory T cells were sorted based on the expression of the congenic marker (CD45.1 or CD45.2) | Congenically marked (CD45.1 or CD45.2) memory T cells were cultured with CD44negCD62LhiCD25-CD4+ (naive) T cells at a 2:1 ratio with anti-CD3/CD28 beads + IL-2 (20 U/ml) + TGFbeta (10 ng/ml) for 48hrs | C57BL/6 | CD44hiCD62L-CD25-CD4+ (Memory) T cells were sorted form spleen and lymph nodes (mesenteric and cervical) |
| GSM335933 | Memory T cells were sorted based on the expression of the congenic marker (CD45.1 or CD45.2) | Congenically marked (CD45.1 or CD45.2) memory T cells were cultured with CD44negCD62LhiCD25-CD4+ (naive) T cells at a 2:1 ratio with anti-CD3/CD28 beads + IL-2 (20 U/ml) + TGFbeta (10 ng/ml) for 48hrs | C57BL/6 | CD44hiCD62L-CD25-CD4+ (Memory) T cells were sorted form spleen and lymph nodes (mesenteric and cervical) |
| GSM335936 | Memory T cells were sorted based on the expression of the congenic marker (CD45.1 or CD45.2) | Congenically marked (CD45.1 or CD45.2) memory T cells were cultured with CD44negCD62LhiCD25-CD4+ (naive) T cells at a 2:1 ratio with anti-CD3/CD28 beads + IL-2 (20 U/ml) + TGFbeta (10 ng/ml) + Retinoic Acid (100nM) for 48hrs | C57BL/6 | CD44hiCD62L-CD25-CD4+ (Memory) T cells were sorted form spleen and lymph nodes (mesenteric and cervical) |
| GSM335936 | Memory T cells were sorted based on the expression of the congenic marker (CD45.1 or CD45.2) | Congenically marked (CD45.1 or CD45.2) memory T cells were cultured with CD44negCD62LhiCD25-CD4+ (naive) T cells at a 2:1 ratio with anti-CD3/CD28 beads + IL-2 (20 U/ml) + TGFbeta (10 ng/ml) + Retinoic Acid (100nM) for 48hrs | C57BL/6 | CD44hiCD62L-CD25-CD4+ (Memory) T cells were sorted form spleen and lymph nodes (mesenteric and cervical) |
| GSM335936 | Memory T cells were sorted based on the expression of the congenic marker (CD45.1 or CD45.2) | Congenically marked (CD45.1 or CD45.2) memory T cells were cultured with CD44negCD62LhiCD25-CD4+ (naive) T cells at a 2:1 ratio with anti-CD3/CD28 beads + IL-2 (20 U/ml) + TGFbeta (10 ng/ml) + Retinoic Acid (100nM) for 48hrs | C57BL/6 | CD44hiCD62L-CD25-CD4+ (Memory) T cells were sorted form spleen and lymph nodes (mesenteric and cervical) |