目的:人循环FoxP3+调节性T细胞亚群表达CD39(cTreg39+)并水解在炎症灶释放的促炎腺嘌呤核苷酸,提供抗炎剂腺苷。甲氨蝶呤(MTX),抑制ATIC,增强腺嘌呤核苷酸的挤出,可能有助于Treg39+细胞控制炎症。因此,我们检查了cTreg39细胞与MTX在早期类风湿关节炎(eRA)中的作用的关系。
方法:通过细胞计数检查了98名未治疗的eRA患者和98名健康对照(HC)的新鲜分离的外周血淋巴细胞。开始MTX后12个月(12m),对82例患者进行了临床重新评估,其中40例重复了细胞计数。在Treg/Tresp共培养物中评估MTX对Treg细胞效力的影响。
结果:基线(0m)cTreg39+细胞频率在eRA中升高到高于HC水平。在12个月时达到低疾病活动度的患者(12m-LDA,DAS28-ESR≤3.2,n=51)的频率明显高于未出现DAS28-ESR的频率(n=31)。获得12m-LDA的0mcTreg39+截止值为42.0%(灵敏度=90.4%/特异性=96.8%)。在12m处,cTreg39+频率不再升高,但其与疾病活动的相关性仍然存在:达到LDA的患者与未达到LDA的患者相比,其仍显著升高.体外,MTX增强了Treg39+细胞效力,但对Treg39-细胞没有影响。
结论:MTX与Treg39+细胞协同作用,基线cTreg39+频率可预测eRA对MTX的反应。此外,eRA中瞬时升高的基线cTreg39+频率可以为提示MTX启动提供一个时隙。
OBJECTIVE: A subset of human circulating FoxP3+ regulatory T cells expresses CD39 (cTreg39+) and hydrolyses pro-inflammatory adenine nucleotides released at inflammatory foci, rendering the anti-inflammatory agent adenosine. Methotrexate (MTX), inhibiting ATIC, enhances the extrusion of adenine nucleotides and may help Treg39+ cells control inflammation. Therefore, we examined the relation of cTreg39+ cells with the effect of MTX in early Rheumatoid Arthritis (eRA).
METHODS: Freshly isolated peripheral blood lymphocytes from 98 untreated eRA patients and 98 healthy controls (HC) were examined by cytometry. Twelve months (12m) after initiating MTX, 82 patients were clinically re-evaluated and cytometry was repeated in 40 of them. The effect of MTX on Treg cell potency was assessed in Treg/Tresp cocultures.
RESULTS: The baseline (0m) cTreg39+ cell frequency was elevated in eRA above HC levels. Patients who reached low disease activity at 12 months (12m-LDA, DAS28-ESR≤ 3.2, n = 51) had presented with a significantly higher 0m cTreg39+ frequency vs those who did not (n = 31). The 0m cTreg39+ cutoff for attaining 12 m-LDA was 42.0% (Sensitivity=90.4%/Specificity=96.8%). At 12m, the cTreg39+ frequency was no longer elevated but its association with disease activity remained: it was still significantly higher in patients who had reached LDA vs those who had not. In vitro, MTX augmented the Treg39+ cell potency but had no effect on Treg39- cells.
CONCLUSIONS: MTX cooperates with Treg39+ cells and the baseline cTreg39+ frequency predicts the response to MTX in eRA. In addition, the transiently elevated baseline cTreg39+ frequency in eRA may provide a slot for prompt MTX initiation.