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High proportion of terminally differentiated regulatory T cells after allogeneic hematopoietic stem cell transplantation

Bone Marrow Transplantation volume 56, pages 1828–1841 (2021)Cite this article

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Abstract

It is now well-established that regulatory T cells (Treg) represent a heterogeneous group of CD4+ T cells. Previous studies have demonstrated that Treg homeostasis was impacted by allogeneic hematopoietic cell transplantation (allo-HCT) and particularly so in patients with chronic graft-versus-host disease (GVHD). Here, we first assessed the ability of various Treg subsets to phosphorylate STAT5 in response to IL-2 or IL-7 stimulation in vitro. We then compared the frequencies of different Treg subtypes in healthy controls as well as in allo-HCT patients with or without chronic GVHD. The highest phosphorylated STAT5 (pSTAT5) signal in response to IL-2 was observed in the CD45RO+CD26−CD39+HLA-DR+ Treg fraction. In contrast, naive Treg were mostly less susceptible to IL-2 stimulation in vitro. Following IL-7 stimulation, most Treg subpopulations upregulated pSTAT5 expression but to a lesser extent than conventional T cells. Compared to healthy controls, allo-HCT patients had lower frequencies of the naive CD45RAbrightCD26+ Treg subpopulation but higher frequencies of the most differentiated memory CD45RO+CD26−CD39+ Treg subpopulations. Further, unbiased analysis revealed that six Treg clusters characterized by high expression of CD25, HLA-DR, and ICOS were significantly more frequent in patients with no or with limited chronic GVHD than in those with moderate/severe chronic GVHD.

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Fig. 1: Impact of IL-2 on Treg subests in vitro.
Fig. 2: Impact of IL-2 on Treg subests in vitro using T-SNE analyses.
Fig. 3: Treg subsets in healthy controls and in patients with or without moderate/severe chronic GVHD.
Fig. 4: T-SNE of Treg subpopulations in healthy controls (n = 20) and in allo-HCT patients without (n = 20) or with (n = 15) moderate/severe chronic GVHD.
Fig. 5: Citrus analyses of Treg (2000/sample) from allo-HCT recipients without (n = 20) or with (n = 15) moderate/severe chronic GVHD.
Fig. 6: Impact of injection of low-dose IL-2 in a patient with severe steroid-refractory chronic GVHD.

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Acknowledgements

This study was supported in part by the European Union’s Horizon 2020 research and innovation programme under Grant agreement No. 643776 (TREGeneration), by the Foundation contre le Cancer (FBC), Belgium, FBC # FAF-C/2016/889 Grant and the National Fund for Scientific Research (FNRS), Belgium, research Grant PDR T006915F and PDR T.0016.20.CR is Televie PhD student and FB is Senior Research Associate at the FNRS, Belgium.

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  1. These authors contributed equally: Caroline Ritacco, Grégory Ehx

Authors and Affiliations

  1. Hematology Research Unit, Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I³, University of Liège, Liège, Belgium

    Caroline Ritacco, Grégory Ehx, Céline Grégoire, Coline Daulne, Sophie Servais, Yves Beguin & Frédéric Baron

  2. Division of Hematology, Department of Medicine, CHU of Liège, Liège, Belgium

    Céline Grégoire, Evelyne Willems, Sophie Servais, Yves Beguin & Frédéric Baron

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Contributions

CR performed the experiments, analyzed the data, and wrote the manuscript; FB wrote the manuscript, designed the study, and interpreted the data; GE performed the experiments, helped in the study design and data interpretation, and edited the manuscript; CG and CD performed the experiments and edited the manuscript; YB helped in the study design, provided clinical data, and edited the manuscript; EW and SS provided clinical data and edited the manuscript. All authors approved the final version of the manuscript.

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Correspondence to Frédéric Baron.

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Conflict of interest

FB has received travel grants and/or speaker honoraria from Celgene, AbbVie, Novartis, Pfizer, and Sanofi. The other authors declare that they have no conflict of interest.

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The study was approved by the Ethics Committee of the CHU of Liège (protocol number TJB1609). All volunteers and patients provided their written informed consent.

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Ritacco, C., Ehx, G., Grégoire, C. et al. High proportion of terminally differentiated regulatory T cells after allogeneic hematopoietic stem cell transplantation. Bone Marrow Transplant 56, 1828–1841 (2021). https://doi.org/10.1038/s41409-021-01221-0

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