Feng Xin, Ramsden Madison K, Negri Julie, Baker Mary Grace, Payne Spencer C, Borish Larry, Steinke John W
Department of Otolaryngology, QiLu Hospital of Shandong University, Jinan, China.
Asthma and Allergic Disease Center, University of Virginia Health System, Charlottesville, Va.
J Allergy Clin Immunol. 2016 Oct;138(4):1089-1097.e3. doi: 10.1016/j.jaci.2016.04.042. Epub 2016 Jun 14.
Aspirin-exacerbated respiratory disease (AERD) differs from aspirin-tolerant disease in part because of eosinophilic tissue infiltration and overexpression of arachidonic acid metabolic pathway components that lead to enhanced secretion of cysteinyl leukotrienes and prostaglandin (PG) D observed constitutively and paradoxically in response to aspirin and other COX inhibitors. We have previously demonstrated the capacity of IFN-γ to drive cysteinyl leukotriene expression and response.
We investigated eosinophils as a source of PGD production in patients with AERD.
Eosinophils were enriched from tissue and peripheral blood obtained from control subjects, patients with aspirin-tolerant disease, and patients with AERD. mRNA was extracted and evaluated for expression of hematopoietic prostaglandin D synthase (hPGDS). Expression of hPGDS protein was confirmed with Western hybridization and immunofluorescence staining. Cells were stimulated with aspirin, and secretion of PGD was quantified. CD34 progenitor cells were isolated and matured into eosinophils in the presence or absence of IFN-γ and hPGDS mRNA, and PGD release was measured.
Gene expression analysis revealed that eosinophils from tissue and blood of patients with AERD display increased levels of hPGDS compared with asthmatic and control samples. Western hybridization confirmed the increase in hPGDS mRNA translated to increased protein expression. Immunofluorescence confirmed mast cells and eosinophils from tissue of patients with AERD and asthma demonstrated hPGDS expression, with higher levels in eosinophils from patients with AERD. Incubation of eosinophils from blood and tissue with aspirin stimulated PGD release. IFN-γ-matured eosinophil progenitors showed enhanced hPGDS expression and increased levels of PGD release at baseline and after aspirin stimulation.
In addition to mast cells, eosinophils represent an important source of PGD in patients with AERD and identify a new target for therapeutic intervention.
阿司匹林诱发的呼吸道疾病（AERD）与阿司匹林耐受型疾病有所不同，部分原因在于嗜酸性粒细胞组织浸润以及花生四烯酸代谢途径成分的过度表达，这会导致半胱氨酰白三烯和前列腺素（PG）D的分泌增强，在对阿司匹林和其他环氧化酶抑制剂产生反应时，这种现象持续且反常地出现。我们之前已经证明了γ干扰素驱动半胱氨酰白三烯表达及反应的能力。
我们研究了嗜酸性粒细胞作为AERD患者中PGD产生来源的情况。
从对照受试者、阿司匹林耐受型疾病患者以及AERD患者获取的组织和外周血中富集嗜酸性粒细胞。提取mRNA并评估造血前列腺素D合酶（hPGDS）的表达。通过蛋白质印迹杂交和免疫荧光染色确认hPGDS蛋白的表达。用阿司匹林刺激细胞，并对PGD的分泌进行定量。分离CD34祖细胞，在有或无γ干扰素和hPGDS mRNA的情况下使其成熟为嗜酸性粒细胞，并测量PGD的释放量。
基因表达分析显示，与哮喘患者和对照样本相比，AERD患者组织和血液中的嗜酸性粒细胞hPGDS水平升高。蛋白质印迹杂交证实hPGDS mRNA的增加转化为蛋白质表达的增加。免疫荧光证实，AERD患者和哮喘患者组织中的肥大细胞和嗜酸性粒细胞显示hPGDS表达，AERD患者嗜酸性粒细胞中的水平更高。用阿司匹林孵育血液和组织中的嗜酸性粒细胞可刺激PGD释放。γ干扰素成熟的嗜酸性粒细胞祖细胞在基线时以及阿司匹林刺激后显示出增强的hPGDS表达和增加的PGD释放水平。
除肥大细胞外，嗜酸性粒细胞是AERD患者中PGD的重要来源，并确定了一个新的治疗干预靶点。
