Proteinase 3 promotes formation of multinucleated giant cells and granuloma-like structures in patients with granulomatosis with polyangiitis.
Proteinase 3 promotes formation of multinucleated giant cells and granuloma-like structures in patients with granulomatosis with polyangiitis.
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DOI：
10.1136/ard-2021-221800
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发表时间：
2023-06
影响因子：
27.4
通讯作者：
中科院分区：
文献类型：
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肉芽肿性多血管炎 (GPA) 和显微镜下多血管炎 (MPA) 是与中性粒细胞和单核细胞中发现的靶向蛋白酶 3 (PR3) 或髓过氧化物酶 (MPO) 的抗中性粒细胞胞浆抗体相关的自身免疫性血管炎。肉芽肿仅见于 GPA，形成于微脓肿部位的多核巨细胞 (MGC) 周围，含有凋亡和坏死的中性粒细胞。由于 GPA 患者中性粒细胞 PR3 表达增加，并且表达 PR3 的凋亡细胞抑制巨噬细胞吞噬作用和细胞清除，因此我们研究了 PR3 在刺激巨细胞和肉芽肿形成中的作用。我们用 PR3 或 MPO 刺激来自 GPA 患者、MPA 患者或健康对照的纯化单核细胞和全外周血单核细胞 (PBMC)，并使用光学、共聚焦和电子显微镜观察 MGC 和肉芽肿样结构的形成，并测量细胞因子的产生。我们研究了单核细胞上 PR3 结合伴侣的表达并测试了其抑制的影响。最后，我们向斑马鱼注射了 PR3，并在新型动物模型中表征了肉芽肿的形成。在体外，PR3 使用来自 GPA 患者的细胞而非来自 MPA 患者的细胞促进单核细胞衍生的 MGC 形成，这依赖于可溶性白介素 6 (IL-6) 以及单核细胞 MAC-1 和蛋白酶激活受体 2，这些物质在 GPA 患者的细胞中过度表达。 PR3刺激的PBMC形成肉芽肿样结构，中央MGC被T细胞包围。 PR3 的这种作用在斑马鱼体内得到了证实，并被氯硝柳胺（一种 IL-6-STAT3 通路抑制剂）所抑制。这些数据为 GPA 中肉芽肿的形成提供了机制基础，并为新的治疗方法提供了理论基础。
Granulomatosis with polyangiitis (GPA) and microscopic polyangiitis (MPA) are autoimmune vasculitides associated with antineutrophil cytoplasm antibodies that target proteinase 3 (PR3) or myeloperoxidase (MPO) found within neutrophils and monocytes. Granulomas are exclusively found in GPA and form around multinucleated giant cells (MGCs), at sites of microabscesses, containing apoptotic and necrotic neutrophils. Since patients with GPA have augmented neutrophil PR3 expression, and PR3-expressing apoptotic cells frustrate macrophage phagocytosis and cellular clearance, we investigated the role of PR3 in stimulating giant cell and granuloma formation. We stimulated purified monocytes and whole peripheral blood mononuclear cells (PBMCs) from patients with GPA, patients with MPA or healthy controls with PR3 or MPO and visualised MGC and granuloma-like structure formation using light, confocal and electron microscopy, as well as measuring the cell cytokine production. We investigated the expression of PR3 binding partners on monocytes and tested the impact of their inhibition. Finally, we injected zebrafish with PR3 and characterised granuloma formation in a novel animal model. In vitro, PR3 promoted monocyte-derived MGC formation using cells from patients with GPA but not from patients with MPA, and this was dependent on soluble interleukin 6 (IL-6), as well as monocyte MAC-1 and protease-activated receptor-2, found to be overexpressed in the cells of patients with GPA. PBMCs stimulated by PR3 formed granuloma-like structures with central MGC surrounded by T cells. This effect of PR3 was confirmed in vivo using zebrafish and was inhibited by niclosamide, a IL-6-STAT3 pathway inhibitor. These data provide a mechanistic basis for granuloma formation in GPA and a rationale for novel therapeutic approaches.
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影响因子：
4.8
作者：
Chen W;Mook RA Jr;Premont RT;Wang J
通讯作者：
Wang J
影响因子：
4.4
作者：
Gabillet, Julie;Millet, Arnaud;Witko-Sarsat, Veronique
通讯作者：
Witko-Sarsat, Veronique
影响因子：
27.4
作者：
Mukhtyar, C.;Lee, R.;Luqmani, R. A.
通讯作者：
Luqmani, R. A.
影响因子：
--
作者：
Exley, AR;Bacon, PA;Adu, D
通讯作者：
Adu, D
影响因子：
3.2
作者：
Anderson, JM
通讯作者：
Anderson, JM