Zinc supplementation augments TGF-β1-dependent regulatory T cell induction
Martina Maywald
Institute of Immunology, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Search for more papers by this authorSteffen K. Meurer
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Search for more papers by this authorRalf Weiskirchen
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Search for more papers by this authorCorresponding Author
Lothar Rink
Institute of Immunology, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Current address: Dr. Lothar Rink, Institute of Immunology, Pauwelsstr. 30, 52074 Aachen, Germany
Correspondence: Dr. Lothar Rink E-mail: [email protected]
Search for more papers by this authorMartina Maywald
Institute of Immunology, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Search for more papers by this authorSteffen K. Meurer
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Search for more papers by this authorRalf Weiskirchen
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Search for more papers by this authorCorresponding Author
Lothar Rink
Institute of Immunology, Faculty of Medicine, RWTH Aachen University Hospital, Aachen, Germany
Current address: Dr. Lothar Rink, Institute of Immunology, Pauwelsstr. 30, 52074 Aachen, Germany
Correspondence: Dr. Lothar Rink E-mail: [email protected]
Search for more papers by this authorAbstract
Scope
Regulatory T cells (Treg) play a pivotal role in immune regulation. For proper immune function, also trace elements such as zinc, and anti-inflammatory cytokines, including transforming growth factor beta 1 (TGF-β1) and interleukin (IL)-10 are indispensable. Hence, in this study the influence of TGF-β1, IL-10, and zinc supplementation on Treg cells differentiation was investigated.
Methods and results
A synergistic effect of a combined zinc and TGF-β1 treatment on Foxp3 expression in peripheral blood mononuclear cells and mixed lymphocyte cultures (MLC) was found by performing Western blot analysis. Additionally, combined treatment causes elevated Smad 2/3 phosphorylation, which plays an important role in Foxp3 expression. This is due to a TGF-β1-mediated increase of intracellular-free zinc measured by zinc probes Fluozin3-AM and ZinPyr-1. Moreover, zinc as well as TGF-β1 treatment caused significantly reduced interferon (IFN)-γ secretion in MLC.
Conclusion
Combined zinc and TGF-β1 treatment provoked an increased Treg cell induction due to a triggered intracellular zinc signal, which in association with an increased Smad 2/3 activation leads to a boosted Foxp3 expression and resulting in an ameliorated allogeneic reaction in MLC. Thus, zinc can be used as a favorable additive to elevate the induction of Treg cells in adverse immune reactions.
Supporting Information
| Filename | Description |
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| mnfr2782-sup-0001-Sup-Mat.docx1,010.8 KB | Supplementary Information Fig. 1. Zinc treatment does not influence the cell viability in MLC experiments. PBMC were adjusted to 2 x 106 cells/mL and remained untreated (white bar) or were pre-incubated with 50 μM zinc (black bar) for 15 min. MLC generation was performed for 5 d. Cell viability was detected by PI staining via FACS analysis and results are displayed. Supplementary Information Fig. 2. Intracellular zinc content in lymphocytes. PBMC were adjusted to 2x106 cells/mL and remained untreated (white bars) or were incubated with 1 ng/mL TGF-β1 or 2 pg/mL IL-10 (grey bars) for indicated time points. Subsequently, intracellular-free zinc was measured by (A, C) Fluozin-3 AM and (B, D) ZinPyr-1 by FACS analysis. Gating was performed on lymphocytes. * indicates a significance of p < 0.05 compared to control, *** indicates a significance of p < 0.001 compared to control (student's t-test). Mean values + SEM are shown. Supplementary Information Figure 3. Intracellular zinc content in monocytes. PBMC were adjusted to 2x106 cells/mL and remained untreated (white bars) or were incubated with 1 ng/mL TGF-β1 or 2 pg/mL IL-10 (grey bars) for indicated time points. Subsequently, intracellular-free zinc was measured by (A, C) Fluozin-3 AM and (B, D) ZinPyr-1 by FACS analysis. Gating was performed on monocytes. Mean values + SEM are shown. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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