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. 2021 May 18;11:10476. doi: 10.1038/s41598-021-88448-5

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Chemokine receptor expression in Treg Populations I–III in HD and MS patients. (A) Representative FACS analysis for CXCR3, CCR6 and CCR7 in CD4⁺ Populations I-V. Expression of chemokine receptors (CCR) was examined in the five Populations of CD4⁺ cells by using FMO control for CCR monoclonal antibody fluorochrome (top row). (B) Representative FACS analysis for CXCR3, CCR6 and CCR7 in Populations I–III within CD4⁺CD25⁺CD127^loTreg. Chemokine receptors (CCR) expression was examined in the three Treg Populations using FMO control for CCR monoclonal antibody fluorochrome (top row). (C) Comparison of chemokine receptor expression in Treg Populations I–III in HD (Ο, n = 20) and MS patients (Δ, n = 31). The proportion of cells expressing CXCR3, CCR6 and CCR7 was examined within CD4⁺CD25⁺CD127^loTreg populations I-III that were identified based on Foxp3 vs. CD45RA expression as outlined in Fig. 2A. Values shown as the percentage of cells expressing respective CCR within each population. Both in HD (n = 19) and in MS patients (n = 35), expression of CXCR3 and CCR6 was higher in Treg Population II compared to Population I (p < 0.0001) or in Population III (p < 0.0001). Population III also had higher expression of CXCR3 and CCR6 compared to Population I (p < 0.0001). CCR7 expression in Treg Population II in both HD (n = 19) and MS (n = 33) was significantly lower compared to Treg Population I (p < 0.0001) or Population III (p < 0.0001). Population III also had significantly lower CCR7 expression compared to Treg Population I in both MS and HD. Approximately half of the MS patients have low CXCR3 and CCR6 expression in Population II. CCR6 expression was lower in Population III in MS compared to HD (p = 0.05).