Dynamic transition of Tregs to cytotoxic phenotype amid systemic inflammation in Graves’ ophthalmopathy
Zhong Liu, … , Zhen Mao, Xian-Chai Lin
Zhong Liu, … , Zhen Mao, Xian-Chai Lin
Published October 4, 2024
Citation Information: JCI Insight. 2024;9(22):e181488. https://doi.org/10.1172/jci.insight.181488.
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Research Article Genetics

Dynamic transition of Tregs to cytotoxic phenotype amid systemic inflammation in Graves’ ophthalmopathy

Abstract

Graves’ disease (GD) is an autoimmune condition that can progress to Graves’ ophthalmopathy (GO), leading to irreversible damage to orbital tissues and potential blindness. The pathogenic mechanism is not fully understood. In this study, we conducted single-cell multi-omics analyses on healthy individuals, patients with GD without GO, newly diagnosed patients with GO, and treated patients with GO. Our findings revealed gradual systemic inflammation during GO progression, marked by overactivation of cytotoxic effector T cell subsets, and expansion of specific T cell receptor clones. Importantly, we observed a decline in the immunosuppressive function of activated Treg (aTreg) accompanied by a cytotoxic phenotypic transition. In vitro experiments revealed that dysfunction and transition of GO-autoreactive Treg were regulated by the yin yang 1 (YY1) upon secondary stimulation of thyroid stimulating hormone receptor (TSHR) under inflammatory conditions. Furthermore, adoptive transfer experiments of the GO mouse model confirmed infiltration of these cytotoxic Treg into the orbital lesion tissues. Notably, these cells were found to upregulate inflammation and promote pathogenic fibrosis of orbital fibroblasts (OFs). Our results reveal the dynamic changes in immune landscape during GO progression and provide direct insights into the instability and phenotypic transition of Treg, offering potential targets for therapeutic intervention and prevention of autoimmune diseases.

Authors

Zhong Liu, Shu-Rui Ke, Zhuo-Xing Shi, Ming Zhou, Li Sun, Qi-Hang Sun, Bing Xiao, Dong-Liang Wang, Yan-Jin Huang, Jin-Shan Lin, Hui-Shi Wang, Qi-Kai Zhang, Cai-Neng Pan, Xuan-Wei Liang, Rong-Xin Chen, Zhen Mao, Xian-Chai Lin

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Figure 6

Pathogenic role of Treg cytotoxic transition to the orbital lesions of GO.

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Pathogenic role of Treg cytotoxic transition to the orbital lesions of G...
(A) Schematic diagram of the EGFP labeled autoreactive Treg adoptive transfer experiment. (B) The proportion of EGFP+KLRC1+ cells within CD4+ cells in the peripheral blood of Healthy and GO mice after adoptive transfer experiment, representing the cytotoxic transition of Treg in vivo (Healthy, n = 3; GO, n = 9). Data are represented as the median IQR. *P < 0.05 by Mann-Whitney U test. (C) Immunofluorescence staining demonstrated the presence of KLRC1+EGFP+ cells in the orbital tissues of GO model mice. (D and E) The major subtypes of orbital tissues (D) and integrated CD4 T cells from PBMCs and tissues (E) based on scRNA-Seq data of Healthy donors and patients with GO; the upper right UMAP plot illustrates the batch effects. (F) The ligand-receptor interactions involving CD4 CTLs significantly altered in GO among 3 LPF subgroups, MYF, and COF. lipofibroblast, LPF; myofibroblast, MYF; conventional orbital fibroblast, COF. Red represents inflammation-regulating ligand-receptor interactions, while green indicates ligand-receptor interactions related to extracellular matrix remodeling. (G) Experimental model of the pathogenic effect of Treg cytotoxic transformation on localized lesions in the orbit. (H) Immunofluorescence showed the expression of specific proteins of OF cells in the coculture system of healthy control and GO model mouse groups; the histograms represent the quantitative statistics of these proteins. Data are represented as the median IQR. **P < 0.001 by Mann-Whitney U test. (I) qPCR histogram showed the expression of specific genes in cells of the coculture system. Each group consists of 3 independent samples (each n = 6). Data are represented as the median IQR. **P < 0.001 by Mann-Whitney U test. All experiments were repeated 3 times.