Evidence for Associations Between Th1/Th17 "Hybrid" Phenotype and Altered Lipometabolism in Very Severe Graves Orbitopathy

Purpose: The purpose of this article is to investigate the characteristics of Th1-cell and Th17-cell lineages for very severe Graves orbitopathy (GO) development.Methods: Flow cytometry was performed with blood samples from GO and Graves disease (GD) patients and healthy controls, to explore effector T-cell phenotypes. Lipidomics was conducted with serum from very severe GO patients before and after glucocorticoid (GC) therapy. Immunohistochemistry and Western blotting were used to examine orbital-infiltrating Th17 cells or in vitro models of Th17 polarization.Results: In GD, Th1 cells predominated in peripheral effector T-cell subsets, whereas in GO, Th17-cell lineage predominated. In moderate-to-severe GO, Th17.1 cells expressed retinoic acid receptor-related orphan receptor-gamma t (ROR gamma t) independently and produced interleukin-17A (IL-17A), whereas in very severe GO, Th17.1 cells co-expressed ROR gamma t and Tbet and produced interferon-gamma (IFN-gamma). Increased IFN-gamma-producing Th17.1 cells positively correlated with GO activity and were associated with the development of very severe GO. Additionally, GC therapy inhibited both Th1-cell and Th17-cell lineages and modulated a lipid panel consisting of 79 serum metabolites. However, in GC-resistant, very severe GO, IFN-gamma-producing Th17.1 cells remained at a high level, correlating with increased serum triglycerides. Further, retro-orbital tissues from GC-resistant, very severe GO were shown to be infiltrated by CXCR3(+) Th17 cells expressing Tbet and STAT4 and rich in triglycerides that promoted Th1 phenotype in Th17 cells in vitro.Conclusions: Our findings address the importance of Th17.1 cells in GO pathogenesis, possibly promoting our understanding of the association between Th17-cell plasticity and disease severity of GO.

目的：本文旨在研究Th1细胞和Th17细胞谱系在极重度格雷夫斯眼病（GO）发展中的特征。 方法：对格雷夫斯眼病和格雷夫斯病（GD）患者以及健康对照的血液样本进行流式细胞术，以探究效应T细胞表型。对极重度格雷夫斯眼病患者糖皮质激素（GC）治疗前后的血清进行脂质组学研究。采用免疫组织化学和蛋白质印迹法检测眼眶浸润的Th17细胞或Th17极化的体外模型。 结果：在GD中，外周效应T细胞亚群以Th1细胞为主，而在GO中，以Th17细胞谱系为主。在中重度GO中，Th17.1细胞独立表达视黄酸受体相关孤儿受体γt（RORγt）并产生白细胞介素 - 17A（IL - 17A），而在极重度GO中，Th17.1细胞共表达RORγt和T - bet并产生干扰素 - γ（IFN - γ）。产生IFN - γ的Th17.1细胞增多与GO活动呈正相关，并与极重度GO的发展有关。此外，GC治疗抑制了Th1细胞和Th17细胞谱系，并调节了由79种血清代谢物组成的脂质组。然而，在GC抵抗的极重度GO中，产生IFN - γ的Th17.1细胞仍处于高水平，与血清甘油三酯升高相关。此外，来自GC抵抗的极重度GO的眶后组织显示有表达T - bet和STAT4的CXCR3⁺ Th17细胞浸润，且富含甘油三酯，甘油三酯在体外促进Th17细胞向Th1表型转化。 结论：我们的研究结果强调了Th17.1细胞在GO发病机制中的重要性，可能有助于我们理解Th17细胞可塑性与GO疾病严重程度之间的关联。