Activation of TGF-β-induced non-Smad signaling pathways during Th17 differentiation
(2015) In Immunology and Cell Biology 93(7). p.662-672- Abstract
Although transforming growth factor-β (TGF-β) has been shown to positively regulate the development of murine T helper type 17 (Th17) cells, which of the intracellular signaling pathways are involved is controversial. We examined Smad-dependent and -independent signaling molecules downstream of the TGF-β receptor (TGFβR) involved in Th17 differentiation of naive murine CD4+ CD62L+ T cells. During Th17 differentiation of wild-type T cells, Smad2/3 was phosphorylated, indicating activation of the canonical Smad pathway. T cells lacking TGFβRII did not differentiate into Th17, whereas T cells treated with a TGFβRI kinase inhibitor (SB-431542) or overexpression of inhibitory Smad7 retained a low amount of Th17... (More)
Although transforming growth factor-β (TGF-β) has been shown to positively regulate the development of murine T helper type 17 (Th17) cells, which of the intracellular signaling pathways are involved is controversial. We examined Smad-dependent and -independent signaling molecules downstream of the TGF-β receptor (TGFβR) involved in Th17 differentiation of naive murine CD4+ CD62L+ T cells. During Th17 differentiation of wild-type T cells, Smad2/3 was phosphorylated, indicating activation of the canonical Smad pathway. T cells lacking TGFβRII did not differentiate into Th17, whereas T cells treated with a TGFβRI kinase inhibitor (SB-431542) or overexpression of inhibitory Smad7 retained a low amount of Th17 polarization despite absent Smad2/3 phosphorylation. Using protein antibody arrays we found an increase of expression and phosphorylation of the following Smad-independent signaling molecules in Th17-polarized wild-type T cells: AKT1(Tyr474), AKT2 (Ser474), ERK1-p44/42 MAPK(Tyr204), mTOR(Thr2446), p38 MAPK(Thr180), Rac1/cdc42(Ser71), SAPK/JNK(Tyr185) and SP1(Thr739). Pharmacological inhibition of AKT/mammalian target of rapamycin (mTOR) signaling with rapamycin or LY294002 decreased Th17 differentiation of wild-type T cells, and completely abolished interleukin-17 production in T cells with overexpression of Smad7. Rapamycin and LY294002 also decreased induced regulatory T cell differentiation, but only had minor additive effects to Smad7 overexpression. Finally, inhibitors of mitogen-activated protein kinase (MAPK) blocked in vitro polarization of Th17 cells. Our data show that Smad-dependent and -independent intracellular pathways contribute to murine Th17 differentiation.
(Less)
- author
- Hasan, Maruf LU ; Neumann, Bernhard ; Haupeltshofer, Steffen LU ; Stahlke, Sarah ; Claudio Fantini, Massimo ; Angstwurm, Klemens ; Bogdahn, Ulrich and Kleiter, Ingo
- publishing date
- 2015-08-12
- type
- Contribution to journal
- publication status
- published
- in
- Immunology and Cell Biology
- volume
- 93
- issue
- 7
- pages
- 662 - 672
- publisher
- Nature Publishing Group
- external identifiers
-
- pmid:25823994
- scopus:84938969665
- ISSN
- 0818-9641
- DOI
- 10.1038/icb.2015.21
- language
- English
- LU publication?
- no
- id
- 542fb272-866c-43ae-93f9-b26b6c074e0a
- date added to LUP
- 2026-02-05 09:24:02
- date last changed
- 2026-02-05 09:25:14
@article{542fb272-866c-43ae-93f9-b26b6c074e0a,
abstract = {{<p>Although transforming growth factor-β (TGF-β) has been shown to positively regulate the development of murine T helper type 17 (Th17) cells, which of the intracellular signaling pathways are involved is controversial. We examined Smad-dependent and -independent signaling molecules downstream of the TGF-β receptor (TGFβR) involved in Th17 differentiation of naive murine CD4<sup>+</sup> CD62L<sup>+</sup> T cells. During Th17 differentiation of wild-type T cells, Smad2/3 was phosphorylated, indicating activation of the canonical Smad pathway. T cells lacking TGFβRII did not differentiate into Th17, whereas T cells treated with a TGFβRI kinase inhibitor (SB-431542) or overexpression of inhibitory Smad7 retained a low amount of Th17 polarization despite absent Smad2/3 phosphorylation. Using protein antibody arrays we found an increase of expression and phosphorylation of the following Smad-independent signaling molecules in Th17-polarized wild-type T cells: AKT1(Tyr474), AKT2 (Ser474), ERK1-p44/42 MAPK(Tyr204), mTOR(Thr2446), p38 MAPK(Thr180), Rac1/cdc42(Ser71), SAPK/JNK(Tyr185) and SP1(Thr739). Pharmacological inhibition of AKT/mammalian target of rapamycin (mTOR) signaling with rapamycin or LY294002 decreased Th17 differentiation of wild-type T cells, and completely abolished interleukin-17 production in T cells with overexpression of Smad7. Rapamycin and LY294002 also decreased induced regulatory T cell differentiation, but only had minor additive effects to Smad7 overexpression. Finally, inhibitors of mitogen-activated protein kinase (MAPK) blocked in vitro polarization of Th17 cells. Our data show that Smad-dependent and -independent intracellular pathways contribute to murine Th17 differentiation.</p>}},
author = {{Hasan, Maruf and Neumann, Bernhard and Haupeltshofer, Steffen and Stahlke, Sarah and Claudio Fantini, Massimo and Angstwurm, Klemens and Bogdahn, Ulrich and Kleiter, Ingo}},
issn = {{0818-9641}},
language = {{eng}},
month = {{08}},
number = {{7}},
pages = {{662--672}},
publisher = {{Nature Publishing Group}},
series = {{Immunology and Cell Biology}},
title = {{Activation of TGF-β-induced non-Smad signaling pathways during Th17 differentiation}},
url = {{http://dx.doi.org/10.1038/icb.2015.21}},
doi = {{10.1038/icb.2015.21}},
volume = {{93}},
year = {{2015}},
}