类风湿关节炎患者血浆细胞外微囊泡中的瓜氨酸化和氨甲酰化蛋白。
Citrullinated and carbamylated proteins in extracellular microvesicles from plasma of patients with rheumatoid arthritis.
机构信息
Department of Clinical Internal, Anesthesiologic and Cardiovascular Sciences, Sapienza University, Rome, Italy.
Department of Experimental Medicine, Sapienza University, Rome, Italy.
出版信息
Rheumatology (Oxford). 2023 Jun 1;62(6):2312-2319. doi: 10.1093/rheumatology/keac598.
OBJECTIVES
To investigate the expression of citrullinated and carbamylated proteins in extracellular microvesicles (EMVs) from RA patients.
METHODS
We enrolled 24 RA naïve for biological therapy and 20 healthy donors (HD), matched for age and sex. For each patient, laboratory and clinical data were recorded and clinical indexes were measured (Clinical Disease Activity Index, Simplified Disease Activity Index, DAS28). EMVs in RA patients and HD were purified from plasma and measured by nanoparticle tracking analysis (NanoSight). Further, EMVs were incubated with anti-citrullinated/carbamylated proteins antibodies and processed by flow cytometry and western blot to evaluate the expression of citrullinated/carbamylated antigens.
RESULTS
NanoSight revealed a significant increase of EMVs in RA compared with HD. Moreover, cytofluorimetric analysis showed a significative higher expression of citrullinated antigens on EMVs' surface in RA than donors, while no substantial difference was found in the expression of carbamylated antigens. These data were confirmed by western blot which identified vimentin, glycolytic enzyme alpha-enolase 1 and collagen type II as the main citrullinated and carbamylated proteins carried by EMVs. Finally, a relevant correlation between the expression of citrullinated antigens and disease activity was found.
CONCLUSIONS
The results of this study suggest an involvement of EMVs in the pathogenesis of RA by inducing autoimmunity.
目的
研究类风湿关节炎（RA）患者细胞外微囊泡（EMVs）中瓜氨酸化和氨甲酰化蛋白的表达。
方法
我们招募了 24 名初治生物治疗的 RA 患者和 20 名健康对照者（HD），年龄和性别相匹配。对每位患者记录实验室和临床数据，并测量临床指标（临床疾病活动指数、简化疾病活动指数、DAS28）。从 RA 患者和 HD 的血浆中纯化 EMVs，并通过纳米颗粒跟踪分析（NanoSight）进行测量。此外，将 EMVs 与抗瓜氨酸化/氨甲酰化蛋白抗体孵育，并用流式细胞术和 Western blot 处理，以评估瓜氨酸化/氨甲酰化抗原的表达。
结果
NanoSight 显示 RA 患者的 EMVs 明显增加。此外，流式细胞分析显示 RA 患者 EMVs 表面瓜氨酸化抗原的表达显著高于供体，而氨甲酰化抗原的表达无明显差异。Western blot 进一步证实了这一点，鉴定出波形蛋白、糖酵解酶α-烯醇化酶 1 和 II 型胶原是 EMVs 携带的主要瓜氨酸化和氨甲酰化蛋白。最后，发现瓜氨酸化抗原的表达与疾病活动度之间存在相关性。
结论
本研究结果提示 EMVs 通过诱导自身免疫参与 RA 的发病机制。
Zotero 插件