Proteinase 3, the Autoantigen in Granulomatosis with Polyangiitis, Associates with Calreticulin on Apoptotic Neutrophils, Impairs Macrophage Phagocytosis, and Promotes Inflammation
Proteinase 3, the Autoantigen in Granulomatosis with Polyangiitis, Associates with Calreticulin on Apoptotic Neutrophils, Impairs Macrophage Phagocytosis, and Promotes Inflammation
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DOI：
10.4049/jimmunol.1200600
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发表时间：
2012-09-01
影响因子：
4.4
通讯作者：
Witko-Sarsat, Veronique
中科院分区：
文献类型：
--
作者：
Gabillet, Julie;Millet, Arnaud;Witko-Sarsat, Veronique
蛋白酶 3 (PR3) 是肉芽肿性多血管炎（系统性血管炎的一种形式）中抗中性粒细胞胞质抗体的靶标。中性粒细胞凋亡后，PR3 与磷脂酰丝氨酸共外化，巨噬细胞吞噬功能受损。钙网蛋白 (CRT) 是一种参与凋亡细胞识别的蛋白质，被发现是一种新的 PR3 伴侣，在凋亡过程中与中性粒细胞质膜上的 PR3 共表达，但在脱粒后则不然。通过共聚焦显微镜和免疫共沉淀在中性粒细胞中证明了 PR3 和 CRT 之间的关联。表面等离子体共振光谱提供了 PR3 与 CRT 球状结构域（但不与其 P 结构域）之间直接相互作用的证据。阻断脂蛋白受体相关蛋白（LRP）（巨噬细胞上的一种 CRT 受体）后，来自健康供体的凋亡中性粒细胞的吞噬作用降低。相反，表达高膜 PR3 水平的肉芽肿性多血管炎患者的中性粒细胞显示出比未受抗 LRP 影响的健康对照者更低的吞噬率，表明 LRP-CRT 通路受到 PR3-CRT 关联的干扰。此外，在体外，人单核细胞来源的巨噬细胞对表达凋亡的PR3的细胞的吞噬作用增强了促炎细胞因子的作用，在体内则通过驻留的小鼠腹腔巨噬细胞增强了促炎细胞因子的作用，并在硫代乙醇酸引发的小鼠巨噬细胞中经LPS攻击后，转移了由凋亡细胞的吞噬作用引发的抗炎反应。因此，凋亡中性粒细胞上表达的膜PR3可能通过影响巨噬细胞的抗炎“重编程”来放大炎症并促进自身免疫。免疫学杂志，2012，189：2574-2583。
Proteinase 3 (PR3) is the target of anti-neutrophil cytoplasm Abs in granulomatosis with polyangiitis, a form of systemic vasculitis. Upon neutrophil apoptosis, PR3 is coexternalized with phosphatidylserine and impaired macrophage phagocytosis. Calreticulin (CRT), a protein involved in apoptotic cell recognition, was found to be a new PR3 partner coexpressed with PR3 on the neutrophil plasma membrane during apoptosis, but not after degranulation. The association between PR3 and CRT was demonstrated in neutrophils by confocal microscopy and coimmunoprecipitation. Evidence for a direct interaction between PR3 and the globular domain of CRT, but not with its P domain, was provided by surface plasmon resonance spectroscopy. Phagocytosis of apoptotic neutrophils from healthy donors was decreased after blocking lipoprotein receptor-related protein (LRP), a CRT receptor on macrophages. In contrast, neutrophils from patients with granulomatosis with polyangiitis expressing high membrane PR3 levels showed a lower rate of phagocytosis than those from healthy controls not affected by anti-LRP, suggesting that the LRP-CRT pathway was disturbed by PR3-CRT association. Moreover, phagocytosis of apoptotic PR3-expressing cells potentiated proinflammatory cytokine in vitro by human monocyte-derived macrophages and in vivo by resident murine peritoneal macrophages, and diverted the anti-inflammatory response triggered by the phagocytosis of apoptotic cells after LPS challenge in thioglycolate-elicited murine macrophages. Therefore, membrane PR3 expressed on apoptotic neutrophils might amplify inflammation and promote autoimmunity by affecting the anti-inflammatory "reprogramming" of macrophages. The Journal of Immunology, 2012, 189: 2574-2583.