低密粒细胞和 NETosis 在成人斯蒂尔病发病机制中的作用。

The role of low density granulocytes and NETosis in the pathogenesis of adult-onset Still's Disease.

机构信息

Department of Immunology and Rheumatology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, and Emergency Medicine Department, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico.

Dept. of Immunology and Rheumatology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, and Laboratory of Clinical Immunology I, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, Mexico.

出版信息

Clin Exp Rheumatol. 2019 Nov-Dec;37 Suppl 121(6):74-82. Epub 2019 Jul 12.

PMID:31365336

Abstract

OBJECTIVES

To analyse the potential contribution of low-density granulocytes (LDGs) and NETosis, as well as the differential protein cargo of neutrophil extracellular traps (NETs), as physiopathogenic mechanisms of adult-onset Still's disease (AOSD).

METHODS

We recruited 30 patients with AOSD according to the Yamaguchi diagnostic criteria. LDGs were addressed by multiparametric flow cytometry as those CD14-, CD15+, CD10+ cells in the peripheral blood mononuclear cells fraction. NETs were quantified by ELISA, immunofluorescence and fluorescence spectrometry. The expression of LL-37 and high mobility group box 1 (HMGB-1) in NETs was measured by immunofluorescence and confocal microscopy. Additionally, normal density neutrophils from healthy controls were stimulated with serum from patients with AOSD and NET induction was assessed by immunofluorescence.

RESULTS

Patients with active disease as well as those with arthritis, cutaneous manifestations and fever had a higher amount of NETs and LDGs. Serum NETs from AOSD patients correlated with the number of swollen joints (r=0.41, p=0.032), absolute number of monocytes (r=0.529, p=0.005). The spontaneous NETs from patients with cutaneous manifestations and fever had higher cargo of HMGB-1 compared with patients in remission.

CONCLUSIONS

LDGs and NETs are increased in patients with active AOSD and correlate with particular clinical features. Patients with cutaneous lesions and fever present a higher cargo of HMGB1 in their spontaneous NETs.

摘要

目的

分析低密粒细胞（LDGs）和 NETosis 的潜在贡献，以及中性粒细胞胞外诱捕网（NETs）的差异蛋白载量，作为成人Still 病（AOSD）的病理生理机制。

方法

我们根据 Yamaguchi 诊断标准招募了 30 名 AOSD 患者。外周血单个核细胞（PBMC）部分中的 CD14-、CD15+、CD10+细胞被多参数流式细胞术确定为 LDGs。通过 ELISA、免疫荧光和荧光光谱法定量 NETs。NETs 中 LL-37 和高迁移率族蛋白 B1（HMGB-1）的表达通过免疫荧光和共聚焦显微镜测量。此外，从健康对照者的正常密度中性粒细胞中提取，并用 AOSD 患者的血清刺激，通过免疫荧光评估 NET 诱导。

结果

活动期疾病患者、关节炎患者、皮肤表现患者和发热患者的 NETs 和 LDGs 数量更高。AOSD 患者的血清 NETs 与肿胀关节数量（r=0.41，p=0.032）、单核细胞绝对值（r=0.529，p=0.005）呈正相关。有皮肤表现和发热的患者的自发 NETs 中 HMGB-1 的含量高于缓解期患者。