Increased monocyte transcription of the proteinase 3 gene in small vessel vasculitis
Increased monocyte transcription of the proteinase 3 gene in small vessel vasculitis
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DOI：
10.1111/j.1365-2249.2005.02819.x
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发表时间：
2005-07-01
影响因子：
4.6
通讯作者：
Segelmark, M
中科院分区：
文献类型：
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作者：
Ohlsson, S;Hellmark, T;Segelmark, M
蛋白酶3（PR 3）是一种多效性和破坏性丝氨酸蛋白酶，也是系统性小血管炎中自身抗体的主要靶点。我们最近发现，稳定缓解的患者循环中PR 3水平升高，与自身抗体滴度、炎症、中性粒细胞脱粒和肾功能无关。在这里，我们探讨增加PR 3基因转录的可能性。从血管炎患者和对照的外周血单核细胞中纯化RNA。通过TaqMan实时聚合酶链反应（PCR）测量特异性mRNA。用细胞因子和脂多糖（LPS）刺激单核细胞样细胞系THP-1和U937以及来自健康对照的人外周血单核细胞不同时间。采用酶联免疫吸附试验（ELISA）测定血浆中PR 3蛋白。22名患者的PR 3 mRNA中位数结果为9.6（1.8-680），而15名健康对照为1（0.1-2.8）。弹性蛋白酶的表达也显着增加，而髓过氧化物酶和白细胞介素-8没有。用肿瘤坏死因子（TNF）-α、干扰素（IFN）-γ或LPS刺激单核细胞不会导致PR 3或弹性蛋白酶转录的任何增加，而白细胞介素（IL）-8转录增加10倍。与健康对照组和系统性红斑狼疮（SLE）患者相比，系统性血管炎患者的循环单核细胞显示PR 3基因转录增加。这可能是重要的发展血管炎。我们的研究结果并不支持细胞因子，抗神经细胞胞浆抗体（ANCA）或免疫抑制药物在血管炎PR 3转录上调的作用。
Proteinase 3 (PR3) is a pleiotropic and destructive serine protease and it is also a major target for autoantibodies in systemic small vessel vasculitis. We have shown recently that patients in stable remission have increased circulating levels of PR3, independent of autoantibody titre, inflammation, neutrophil degranulation and renal function. Here we explore the possibility of increased PR3 gene transcription. RNA was purified from peripheral blood monocytes from vasculitis patients and controls. Specific mRNA was measured by TaqMan real-time polymerase chain reaction (PCR). The monocyte-like cell lines THP-1 and U937 and human peripheral blod monocytes from healthy controls were stimulated with cytokines and lipopolysaccharide (LPS) for different time periods. PR3 protein was measured in plasma with enzyme-linked immunosorbent assay (ELISA). The median result for PR3 mRNA was 9.6 (1.8-680) for 22 patients, compared to 1 (0.1-2.8) for the 15 healthy controls. Elastase expression was also significantly increased, whereas myeloperoxidase and interleukin-8 were not. Stimulation of monocytes with tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma or LPS did not result in any increase of PR3 or elastase transcription, whereas interleukin (IL)-8 transcription was increased 10-fold. Circulating monocytes from patients with systemic vasculitis display increased PR3 gene transcription compared to healthy controls and patients with sytemic lupus erythematosus (SLE). This may be important for the development of vasculitis. Our results do not favour a role for cytokines, antineutrophil cytoplasmic antibodies (ANCA) or immunosuppressive medication in the upregulation of PR3 transcription in vasculitis.