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. 2022 Jan 17;12:850. doi: 10.1038/s41598-021-04442-x

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022

Open AccessThis 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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Changes in Th1 and Treg cell populations and function are largely due to intrinsic loss of Irgm1 expression. (a-b) Splenocytes were isolated from LckCre⁺Irgm1^fl/fl cKO mice and LckCre⁻Irgm1^fl/fl littermate controls, stained and analyzed by flow cytometry for CD4 and CD8 cells (a), CD4⁺IFNγ⁺ and CD4⁺IL-17⁺ (b). (c) CD4⁺ T cells were isolated and activated for 48 h with plate bound anti-CD3 and anti-CD28 antibodies, after which the media was collected and IFNγ and IL-17A were measured by ELISA. (d) In vitro differentiated Treg cells from LckCre⁺Irgm1^fl/fl cKO mice and LckCre⁻Irgm1^fl/fl littermate controls were co-cultured for three days with WT CD8⁺ T cells stained with CellTrace violet. Treg suppressive capacity was analyzed by the ability to reduce proliferation of CD8⁺ T cells, as measured by dilution of fluorescent dye flow cytometrically (The CD8:Treg ratio for the data shown is 2:1). (a, b) data pooled from two independent experiments (n = 6), (c, d) data representative of two independent experiments (n = 3–6); error bars represent ± SEM. Mann Whitney test or T-test with Welch’s correction was used to compare groups depending on the normality of the distribution as judged by the Shapiro–Wilk test.*p < 0.05, **p < 0.01, ***p < 0.001.