健康受试者和类风湿性关节炎患者中性粒细胞亚群氧化反应的差异。

Differences in oxidative response of subpopulations of neutrophils from healthy subjects and patients with rheumatoid arthritis.

作者信息

Eggleton P, Wang L, Penhallow J, Crawford N, Brown K A

机构信息

Department of Biochemistry, University of Oxford, United Kingdom.

出版信息

Ann Rheum Dis. 1995 Nov;54(11):916-23. doi: 10.1136/ard.54.11.916.

DOI:10.1136/ard.54.11.916

PMID:7492242

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1010043/

Abstract

OBJECTIVES

To determine whether blood neutrophils from healthy individuals and blood and synovial fluid neutrophils from patients with rheumatoid arthritis (RA) responded differently to priming agonists and stimuli of the oxidative burst and, if so, whether this was a property of a subpopulation of neutrophils.

METHODS

Continuous flow electrophoresis was used to separate neutrophils into subpopulations based upon quantitative differences in net negative surface charge. The generation of superoxide anion (O2-) was used as a measure of oxidative activity using 10(-7) mol/l N-formyl-methionylleucyl-phenylalanine (FMLP) as the stimulating agonist and 10(-8) mol/l platelet activating factor (PAF) as the priming agent.

RESULTS

The production of O2- by blood and synovial fluid neutrophils from RA patients in response to FMLP was greater than that observed with control blood neutrophils (p < 0.001). Priming of normal blood neutrophils with PAF increased their FMLP induced oxidative burst (p < 0.001), but PAF treatment had no effect on rheumatoid neutrophils. Neutrophils from synovial fluid of RA patients were less electronegative than paired blood samples and exposure of blood neutrophils to FMLP but not PAF reduced their surface charge. Continuous flow electrophoresis isolated three neutrophil subpopulations: cells of least surface electronegativity were ascribed to pool P1 and cells of greatest surface electro-negativity to P3. Normal blood neutrophils from P3, but not P1, showed increased oxidative activity after PAF priming (twofold increase; p < 0.01), whereas the responsiveness of rheumatoid blood and synovial fluid neutrophils from P1 and P3 was not modified by PAF treatment under the same conditions.

CONCLUSION

It is suggested that most of the circulating neutrophils in RA are already in a state of readiness to generate O2- upon activation by an inflammatory stimulus. This is in contrast to normal blood neutrophils, which have both responsive and non-responsive subpopulations with respect to priming agonists.

摘要

目的

确定健康个体的血液中性粒细胞以及类风湿关节炎（RA）患者的血液和滑液中性粒细胞对引发激动剂和氧化爆发刺激的反应是否不同，若不同，这是否是中性粒细胞亚群的特性。

方法

采用连续流动电泳根据净负表面电荷的定量差异将中性粒细胞分离成亚群。使用10⁻⁷mol/L的N-甲酰甲硫氨酰亮氨酰苯丙氨酸（FMLP）作为刺激激动剂，10⁻⁸mol/L的血小板活化因子（PAF）作为引发剂，通过超氧阴离子（O₂⁻）的生成来衡量氧化活性。

结果

RA患者血液和滑液中性粒细胞对FMLP反应产生的O₂⁻ 量大于对照血液中性粒细胞（p < 0.001）。用PAF引发正常血液中性粒细胞可增加其FMLP诱导的氧化爆发（p < 0.001），但PAF处理对类风湿中性粒细胞无影响。RA患者滑液中的中性粒细胞比配对血液样本的电负性更低，血液中性粒细胞暴露于FMLP而非PAF会降低其表面电荷。连续流动电泳分离出三个中性粒细胞亚群：表面电负性最小的细胞归为P1池，表面电负性最大的细胞归为P3池。PAF引发后，P3池的正常血液中性粒细胞而非P1池的，氧化活性增加（增加两倍；p < 0.01），而在相同条件下，PAF处理并未改变P1池和P3池的类风湿血液和滑液中性粒细胞的反应性。