METHODS: Peripheral blood samples from 37 healthy controls and 44 pSS patients were collected. The publicly available single-cell RNA-Seq data from pSS patient PBMCs were utilized to analyse KLRB1 expression in T cells. KLRB1-expressing T lymphocyte subset proportions in pSS patients and healthy controls were determined by flow cytometry. CD25, Ki-67, cytokine secretion, and chemokine receptor expression in CD4+ KLRB1+ T cells were detected and compared with those in CD4+ KLRB1- T cells. Correlation analysis was conducted between KLRB1-related T-cell subsets and clinical indicators. ROC curves were generated to explore the diagnostic potential of KLRB1 for pSS.
RESULTS: KLRB1 was significantly upregulated following T-cell activation, and Ki-67 and CD25 expression was significantly greater in CD4+ KLRB1+ T cells than in CD4+ KLRB1- T cells. KLRB1+ CD4+ T cells exhibited greater IL-17A, IL-21, IL-22, and IFN-γ secretion upon stimulation, and there were significantly greater proportions of CCR5+, CCR2+, CX3CR1+, CCR6+, and CXCR3+ cells among CD4+ KLRB1+ T cells than among CD4+ KLRB1- T cells. Compared with that in HCs, KLRB1 expression in CD4+ T cells was markedly elevated in pSS patients and significantly correlated with clinical disease indicators.
CONCLUSIONS: KLRB1 is a characteristic molecule of the CD4+ T-cell activation phenotype. The increased expression of KLRB1 in the CD4+ T cells of pSS patients suggests its potential involvement in the pathogenesis of pSS and its utility as an auxiliary diagnostic marker for pSS.
方法:收集37名健康对照和44名pSS患者的外周血样本。来自pSS患者PBMC的公开可用的单细胞RNA-Seq数据用于分析T细胞中的KLRB1表达。通过流式细胞术确定pSS患者和健康对照组中表达KLRB1的T淋巴细胞亚群比例。CD25,Ki-67,细胞因子分泌,检测CD4+KLRB1+T细胞趋化因子受体的表达,并与CD4+KLRB1-T细胞进行比较。对KLRB1相关T细胞亚群与临床指标进行相关性分析。产生ROC曲线以探索KLRB1对pSS的诊断潜力。
结果:KLRB1在T细胞活化后显著上调,CD4+KLRB1+T细胞中Ki-67和CD25表达显著高于CD4+KLRB1-T细胞。KLRB1+CD4+T细胞表现出更高的IL-17A,刺激后IL-21、IL-22和IFN-γ分泌,CCR5+的比例明显更高,CCR2+,CX3CR1+,CCR6+,CD4+KLRB1+T细胞中的CXCR3+细胞高于CD4+KLRB1-T细胞中的CXCR3+细胞。与HC相比,pSS患者CD4+T细胞中KLRB1的表达明显升高,与临床疾病指标显著相关。
结论:KLRB1是CD4+T细胞活化表型的特征性分子。KLRB1在pSS患者的CD4T细胞中的表达增加表明其可能参与pSS的发病机理及其作为pSS辅助诊断标志物的用途。