Recirculating IL-1R2+ Tregs fine-tune intrathymic Treg development under inflammatory conditions

Abstract

The vast majority of Foxp3+ regulatory T cells (Tregs) are generated in the thymus, and several factors, such as cytokines and unique thymic antigen-presenting cells, are known to contribute to the development of these thymus-derived Tregs (tTregs). Here, we report the existence of a specific subset of Foxp3+ Tregs within the thymus that is characterized by the expression of IL-1R2, which is a decoy receptor for the inflammatory cytokine IL-1. Detailed flow cytometric analysis of the thymocytes from Foxp3hCD2xRAG1GFP reporter mice revealed that the IL-1R2+ Tregs are mainly RAG1GFP– and CCR6+CCR7, demonstrating that these Tregs are recirculating cells entering the thymus from the periphery and that they have an activated phenotype. In the spleen, the majority of IL-1R2+ Tregs express neuropilin-1 (Nrp-1) and Helios, suggesting a thymic origin for these Tregs. Interestingly, among all tissues studied, the highest frequency of IL-1R2+ Tregs was observed in the thymus, indicating preferential recruitment of this Treg subset by the thymus. Using fetal thymic organ cultures (FTOCs), we demonstrated that increased concentrations of exogenous IL-1β blocked intrathymic Treg development, resulting in a decreased frequency of CD25+Foxp3+ tTregs and an accumulation of CD25+Foxp3 Treg precursors. Interestingly, the addition of IL-1R2+ Tregs, but not IL-1R2 Tregs, to reaggregated thymic organ cultures (RTOCs) abrogated the IL-1β-mediated blockade, demonstrating that these recirculating IL-1R2+ Tregs can quench IL-1 signaling in the thymus and thereby maintain thymic Treg development even under inflammatory conditions.

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Fig. 1: Phenotyping IL-1R2+Foxp3+ Tregs in the thymus.
Fig. 2: Phenotyping IL-1R2+Foxp3+ Tregs in the spleen.
Fig. 3: Expression level of IL-1R2 on tissue-resident Tregs.
Fig. 4: Expression of IL-1R1 and its ligands, IL-1α and IL-1β, in the thymus.
Fig. 5: Exogenous IL-1β affects tTreg development in the FTOCs.
Fig. 6: Normal tTreg generation despite increased intrathymic IL-1β levels after CFA treatment.
Fig. 7: IL-1R2+Foxp3+ Tregs fine-tune tTreg development in the RTOCs under inflammatory conditions.

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Acknowledgements

We thank Marina Wuttke and Lothar Groebe for their technical and cell sorting support. This work was supported by the CRC 738 (to J.H.), CRC/TR 128 (to A.W.) and CRC/TRR 221 (to M.F.) of the German Research Foundation and by grants from the European Research Council (ERC-CoG, #648145 REGiREG) to M.F. Y.E. was supported by Ph.D scholarship program no. 57129429 of the German Academic Exchange Service (DAAD).

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E.N., Y.E., S.H., M.D., C.S. and I.A.M. performed the experiments and interpreted the data. A.W., C.F. and M.F. interpreted the data. E.N. and J.H. designed the research, interpreted the data, and wrote the paper.

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Correspondence to Jochen Huehn.

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Nikolouli, E., Elfaki, Y., Herppich, S. et al. Recirculating IL-1R2+ Tregs fine-tune intrathymic Treg development under inflammatory conditions. Cell Mol Immunol (2020). https://doi.org/10.1038/s41423-019-0352-8

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Keywords

  • Thymus
  • Treg development
  • Inflammation
  • IL-1 system