Zhang Dongdong, Alip Mihribangvl, Chen Hongzhen, Wu Dan, Zhu Huimin, Han Yichen, Yuan Xinran, Feng Xuebing, Sun Lingyun, Wang Dandan
Department of Rheumatology and Immunology, Affiliated Hospital of Medical School, Nanjing Drum Tower Hospital, Nanjing University. Nanjing, Jiangsu, 210008, China.
Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine Nanjing, Jiangsu, 210008, China.
Clin Rheumatol. 2024 May;43(5):1623-1634. doi: 10.1007/s10067-024-06920-9. Epub 2024 Mar 4.
To construct a molecular immune map of patients with systemic sclerosis (SSc) by mass flow cytometry, and compare the number and molecular expression of double-negative T (DNT) cell subsets between patients and healthy controls (HC).
Peripheral blood mononuclear cells (PBMCs) were extracted from the peripheral blood of 17 SSc patients and 9 HC. A 42-channel panel was set up to perform mass cytometry by time of flight (CyTOF) analysis for DNT subgroups. Flow cytometry was used to validate subpopulation functions. The clinical data of patients were collected for correlation analysis.
Compared with HC, the number of total DNT cells decreased in SSc patients. Six DNT subsets were obtained from CyTOF analysis, in which the proportion of cluster1 increased, while the proportion of cluster3 decreased. Further analysis revealed that cluster1 was characterized by high expression of CD28 and CCR7, and cluster3 was characterized by high expression of CD28 and CCR5. After in vitro stimulation, cluster1 secreted more IL-4 and cluster3 secreted more IL-10 in SSc patients compared to HC. Clinical correlation analysis suggested that cluster1 may play a pathogenic role while cluster3 may play a protective role in SSc. ROC curve analysis further revealed that cluster3 may be a potential indicator for determining disease activity in SSc patients.
We found a new CCR5CD28 DNT cell subset, which played a protective role in the pathogenesis of SSc. Key Points • The number of DNT cells decreased in SSc patients' peripheral blood. • DNT cells do not infiltrate in the skin but secrete cytokines to participate in the pathogenesis of SSc. • A CCR5CD28 DNT cell population may play a protective role in SSc.
通过质谱流式细胞术构建系统性硬化症（SSc）患者的分子免疫图谱，并比较患者与健康对照（HC）之间双阴性T（DNT）细胞亚群的数量和分子表达。
从17例SSc患者和9例HC的外周血中提取外周血单个核细胞（PBMC）。设置一个42通道的面板，通过飞行时间质谱流式细胞术（CyTOF）分析DNT亚群。采用流式细胞术验证亚群功能。收集患者的临床资料进行相关性分析。
与HC相比，SSc患者中总DNT细胞数量减少。通过CyTOF分析获得了6个DNT亚群，其中cluster1的比例增加，而cluster3的比例降低。进一步分析显示，cluster1的特征是CD28和CCR7高表达，cluster3的特征是CD28和CCR5高表达。体外刺激后，与HC相比，SSc患者中cluster1分泌更多的IL-4，cluster3分泌更多的IL-10。临床相关性分析表明，cluster1可能在SSc中起致病作用，而cluster3可能起保护作用。ROC曲线分析进一步显示，cluster3可能是确定SSc患者疾病活动度的潜在指标。
我们发现了一个新的CCR5CD28 DNT细胞亚群，其在SSc发病机制中起保护作用。要点•SSc患者外周血中DNT细胞数量减少。•DNT细胞不浸润皮肤，但分泌细胞因子参与SSc的发病机制。•一个CCR5CD28 DNT细胞群体可能在SSc中起保护作用。
