Localization of eosinophil-derived neurotoxin and eosinophil cationic protein in neutrophilic leukocytes.
Localization of eosinophil-derived neurotoxin and eosinophil cationic protein in neutrophilic leukocytes.
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嗜酸性粒细胞衍生的神经毒素和嗜酸性粒细胞阳离子蛋白在中性粒细胞中的定位。
DOI：
10.1159/000024091
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发表时间：
1999
影响因子：
2.8
通讯作者：
Leiferman,KM
中科院分区：
文献类型：
--
作者：
Sur,S;Glitz,DG;Kita,H;Kujawa,SM;Peterson,EA;Weiler,DA;Kephart,GM;Wagner,JM;George,TJ;Gleich,GJ;Leiferman,KM
嗜酸性粒细胞衍生神经毒素（EDN）和嗜酸性粒细胞阳离子蛋白（ECP）是嗜酸性粒细胞中独特的颗粒相关蛋白[1,2,3,4,5,6,7]。本研究的目的是确定EDN和ECP是否存在于成熟的中性粒细胞中。通过间接免疫荧光，嗜酸性粒细胞和中性粒细胞都被EDN和ECP抗体染色（图1）。用EDN抗体染色免疫金标记的外周血细胞电镜检查显示，EDN定位于中性粒细胞颗粒（图2）。嗜酸性粒细胞污染小于0.1%的纯化中性粒细胞裂解物（表1）含有EDN（112±4 ng/ 10.6个细胞）和ECP（163±2 ng/ 10.6个细胞），而嗜酸性粒细胞主要碱性蛋白在中性粒细胞制剂中检测不到，但在嗜酸性粒细胞制剂中检测到（表2）。最后，通过逆转录聚合酶链反应（RT-PCR）在中性粒细胞裂解物中检测EDN mRNA（图3），包括高度纯化的中性粒细胞（0.001%嗜酸性粒细胞污染，图4）。根据这些观察和之前的报道，EDN mRNA在嗜中性粒细胞分化的HL-60细胞中表达，并且EDN mRNA存在于成熟的中性粒细胞[7]中，我们得出结论，中性粒细胞中存在与EDN和ECP相同或免疫交叉反应的蛋白质，EDN在中性粒细胞颗粒[8]中合成并定位。因此，在将EDN和ECP的存在解释为人类疾病中嗜酸性粒细胞相关炎症[9]的特异性标志物时，必须谨慎。
Eosinophil–derived neurotoxin (EDN) and eosinophil cationic protein (ECP) are distinctive granule–associated proteins in eosinophils [1, 2, 3, 4, 5, 6, 7]. The purpose of this investigation was to determine whether EDN and ECP are present in mature neutrophils. By indirect immunofluorescence, both eosinophils and neutrophils stained with antibodies to EDN and ECP (fig. 1). Electron–microscopic examination of immunogold–labeled peripheral blood cells stained with EDN antibody showed that EDN was localized to neutrophil granules (fig. 2). Lysates of purified neutrophils with less than 0.1% eosinophil contamination (table 1) contained EDN, 112±4 ng/10 6 cells, and ECP, 163±2 ng/10 6 cells, whereas eosinophil major basic protein was not detectable in the neutrophil preparations but was detectable in eosinophil preparations (table 2). Finally, EDN mRNA was detected in neutrophil lysates (fig. 3), including highly purified neutrophils (0.001% eosinophil contamination, fig. 4) by the reverse transcription–polymerase chain reaction (RT–PCR). Based on these observations and the previous reports that EDN mRNA is expressed in HL–60 cells committed toward neutrophilic differentiation and that EDN mRNA is present in mature neutrophils [7], we conclude that proteins either identical to or immunologically cross–reactive with EDN and ECP are present in neutrophils and that EDN is synthesized in and localized to neutrophil granules [8]. Thus, caution must be exercised in interpreting the presence of EDN and ECP as specific markers of eosinophil–associated inflammation [9] in human disease.