Peters M S, Rodriguez M, Gleich G J
Lab Invest. 1986 Jun;54(6):656-62.
By utilizing the colloidal gold particle technique, we localized eosinophil granule major basic protein, eosinophil cationic protein (ECP), and eosinophil-derived neurotoxin (EDN) in human nasal polyp sections by immunoelectron microscopy. Sections stained with affinity chromatography purified rabbit anti-human major basic protein, and subsequently with gold colloidal particle-goat anti-rabbit IgG, showed gold particles predominantly within granule cores, and not within other eosinophil organelles, plasma cells, mast cells, lymphocytes, or neutrophils. Sections stained with anti-ECP or anti-EDN showed gold particles concentrated over the granule matrix with fewer particles centrally. Control sections treated with preimmunization sera showed no staining of cells or organelles. These results verify the localization of major basic protein to the crystalloid core of the human eosinophil granule and show that ECP and EDN reside in the granule matrix. This technique provides a means of accurately locating the sites of major basic protein, ECP, and EDN deposition and thus of identifying eosinophil degranulation patterns in human disease.
通过运用胶体金颗粒技术,我们采用免疫电子显微镜在人鼻息肉切片中定位了嗜酸性粒细胞颗粒主要碱性蛋白、嗜酸性粒细胞阳离子蛋白(ECP)和嗜酸性粒细胞衍生神经毒素(EDN)。用亲和层析纯化的兔抗人主要碱性蛋白染色的切片,随后用胶体金颗粒-羊抗兔IgG染色,结果显示金颗粒主要位于颗粒核心内,而非其他嗜酸性粒细胞细胞器、浆细胞、肥大细胞、淋巴细胞或中性粒细胞内。用抗ECP或抗EDN染色的切片显示金颗粒集中在颗粒基质上,核心处颗粒较少。用免疫前血清处理的对照切片未显示细胞或细胞器染色。这些结果证实了主要碱性蛋白定位于人嗜酸性粒细胞颗粒的晶体核心,并表明ECP和EDN存在于颗粒基质中。该技术提供了一种准确定位主要碱性蛋白、ECP和EDN沉积部位的方法,从而可识别人类疾病中的嗜酸性粒细胞脱颗粒模式。