蕈样肉芽肿患者瘙痒与趋化因子CCL17相关性研究

Correlations between pruritus and CC chemokine ligand 17 in patients with mycosis fungoides

目的:寻找参与蕈样肉芽肿(MF)患者瘙痒发生的相关分子。方法:纳入2009年10月至2021年8月于北京大学第一医院就诊的522例MF患者作为研究对象,统计瘙痒发生率。根据是否有瘙痒症状对患者进行分组,分析其中49例患者皮损活检组织的RNA测序数据,寻找有瘙痒症状与无瘙痒症状患者的差异表达基因;使用酶联免疫吸附试验和免疫组织化学技术分别检测88例MF患者血清和81例MF患者组织CC趋化因子17(CCL17)蛋白表达量;使用流式细胞仪检测46例患者外周血T淋巴细胞活化和分化标记,寻找与瘙痒症状相关的外周血淋巴细胞亚群。使用χ^(2)检验、Mann-Whitney U检验、Spearman相关性检验进行统计分析。结果:522例患者中,男305例,女217例;早期MF 347例,进展期MF 175例。MF患者瘙痒发生率为67.2%(351例),进展期患者瘙痒发生率(81.7%,143/175)较早期患者(59.9%,208/347)上升(χ^(2)=25.03,P<0.001)。RNA测序结果显示,瘙痒MF患者CCL17 mRNA表达量高于无瘙痒MF患者(差异表达倍数=10.09,P<0.001)。瘙痒患者血清CCL17浓度[1017.05(377.12,4831.80)pg/ml]较无瘙痒患者[361.66(180.47,500.08)pg/ml]上升(Z=-4.57,P<0.001),且与瘙痒评分呈正相关(r=0.57,P=0.010)。在早期和进展期MF患者中,瘙痒患者血清CCL17浓度均高于无瘙痒患者(Z=-3.68,P<0.001;Z=-2.54,P=0.011)。瘙痒患者与无瘙痒患者CCL17免疫组化相对定量评分差异无统计学意义(Z=-1.84,P=0.066)。瘙痒患者CD3+CD4+CD26-CCR4+恶性T细胞百分比高于无瘙痒MF患者(Z=-2.03,P=0.043),且与血清CCL17浓度呈正相关(r=0.49,P<0.001)。结论:MF瘙痒患者皮损组织CCL17mRNA和血清CCL17浓度上升,CCL17与MF患者瘙痒发生相关,可能通过招募CD3+CD4+CD26-CCR4+恶性T细胞参与瘙痒发生。

Objective To investigate molecules involved in the occurrence of pruritus in patients with mycosis fungoides(MF).Methods Totally,522 patients with MF were enrolled from Peking University First Hospital from October 2009 to August 2021,and the incidence of pruritus was calculated.The patients were grouped according to whether they suffered from pruritus or not.RNA sequencing data on biopsied skin lesions of 49 patients were analyzed to identify differentially expressed genes between patients with pruritus and those without;enzyme-linked immunosorbent assay and immunohistochemical techniques were performed to determine the protein expression of CC chemokine ligand 17(CCL17)in serum samples from 88 MF patients,and in tissue samples from 81 MF patients,respectively;flow cytometry was conducted to detect markers for T lymphocyte activation and differentiation in peripheral blood samples from 46 MF patients to identify peripheral blood lymphocyte subsets associated with pruritus.Statistical analysis was carried out by using chi-square test,Mann-Whitney U test,and Spearman correlation analysis.Results Among the 522 patients with MF,305 were males and 217 were females;347 were diagnosed with early-stage MF,and 175 with advanced MF.The incidence of pruritus was 67.2%(351/522)in the patients with MF,and significantly higher in the patients with advanced MF(81.7%,143/175)than in those with early-stage MF(59.9%,208/347;χ^(2)=25.03,P<0.001).RNA sequencing showed that CCL17 mRNA expression was significantly higher in the MF patients with pruritus than in those without(fold change=10.09,P<0.001).The serum CCL17 concentration was significantly elevated in the patients with pruritus(1017.05[377.12,4831.80]pg/ml)compared with those without(361.66[180.47,500.08]pg/ml;Z=-4.57,P<0.001),and correlated with pruritus scores(r=0.57,P=0.010).In both early and advanced stages of MF,the serum CCL17 concentration was significantly higher in the patients with pruritus than in those without(Z=-3.68,P<0.001;Z=-2.54,P=0.011,respectively).Immunohistochemical staining revealed that there was no significant difference in the relative quantification value of CCL17 between the patients with pruritus and those without(Z=-1.84,P=0.066).The percentage of CD3+CD4+CD26-CCR4+malignant T cells significantly increased in the MF patients with pruritus than in those without(Z=-2.03,P=0.043),and was positively correlated with serum CCL17 concentrations(r=0.49,P<0.001).Conclusions Both CCL17 mRNA expression in lesional tissues and serum CCL17 concentrations increased in MF patients with pruritus,and CCL17 was associated with the occurrence of pruritus.CCL17 may be involved in the occurrence of pruritus through the recruitment of CD3+CD4+CD26-CCR4+malignant T cells.