Karawajczyk M, Sevéus L, Garcia R, Björnsson E, Peterson C G, Roomans G M, Venge P
Department of Medical Sciences, Clinical Chemistry, University of Uppsala, Sweden.
Am J Respir Cell Mol Biol. 2000 Oct;23(4):521-9. doi: 10.1165/ajrcmb.23.4.4025.
The variability of serum and plasma levels of eosinophil granule proteins in different clinical conditions, interpreted as the result of different patterns of cytokine priming, suggests a selective mobilization of granule proteins. Inasmuch as piecemeal degranulation (PM) is the mechanism proposed for the differential release of eosinophil granule proteins, we decided to investigate whether blood eosinophils from allergic subjects show characteristics of PM during natural allergen challenge. Eosinophils from three birch-sensitive subjects were studied before and during the pollen season. Electron microscopy analysis showed that during the season, eosinophils presented morphologic features of PM. By immunogold labeling, eosinophil cationic protein (ECP) was detected not only in normal specific granules but also in the cytoplasm, in the vicinity of partially lucent specific granules. These results were confirmed by subcellular fractionation, where the amount of ECP associated with compartments containing small vesicles increased 2-fold during the pollen season. A study of the distribution of ECP, eosinophil peroxidase, and hexosaminidase in eosinophils of different densities showed that the profile of each of these proteins differed depending on cell density. All of these proteins decreased in the specific granule of hypodense cells and increased in other cell compartments. We conclude that allergen exposure causes PM of the peripheral blood eosinophils of allergic subjects, and that the density of these cells reflects the degree of degranulation. Our results provide novel information for the understanding of the selective mobilization of granule proteins into the circulation.
在不同临床状况下,血清和血浆中嗜酸性粒细胞颗粒蛋白水平的变异性,被解释为细胞因子启动模式不同的结果,这表明颗粒蛋白存在选择性动员。鉴于逐片脱颗粒(PM)是被提出用于解释嗜酸性粒细胞颗粒蛋白差异释放的机制,我们决定研究过敏受试者的血液嗜酸性粒细胞在自然过敏原激发过程中是否表现出PM的特征。对三名桦树过敏受试者在花粉季节前后的嗜酸性粒细胞进行了研究。电子显微镜分析显示,在花粉季节期间,嗜酸性粒细胞呈现出PM的形态学特征。通过免疫金标记,不仅在正常的特异性颗粒中检测到嗜酸性粒细胞阳离子蛋白(ECP),而且在细胞质中、部分透亮的特异性颗粒附近也检测到了ECP。亚细胞分级分离证实了这些结果,在花粉季节期间,与含有小泡的区室相关的ECP量增加了2倍。对不同密度嗜酸性粒细胞中ECP、嗜酸性粒细胞过氧化物酶和己糖胺酶分布的研究表明,这些蛋白质中每种的分布情况因细胞密度而异。所有这些蛋白质在低密度细胞的特异性颗粒中减少,而在其他细胞区室中增加。我们得出结论,过敏原暴露会导致过敏受试者外周血嗜酸性粒细胞发生PM,并且这些细胞的密度反映了脱颗粒程度。我们的结果为理解颗粒蛋白向循环中的选择性动员提供了新信息。