Sevéus L, Amin K, Peterson C G, Roomans G M, Venge P
Pharmacia and Upjohn Diagnostics, Uppsala, Sweden.
Histochem Cell Biol. 1997 May;107(5):423-32. doi: 10.1007/s004180050129.
The neutrophilic granulocyte is a cytotoxic and potentially tissue-injuring cell participating in the destructive processes and symptoms seen in a variety of inflammatory diseases. Sensitive immunoassays have been introduced to measure the levels of specific secretory proteins of various inflammatory cells in blood and other body fluids. The aim has been to develop highly specific markers for each cell type. The results obtained by immunoassay have indicated that human neutrophil lipocalin (HNL) is a protein unique to the neutrophil. The present study investigated the specificity of HNL as a neutrophil marker in peripheral blood and lung tissue by using flow cytometry and immunocytochemistry. Flow cytometry and immunocytochemistry on peripheral blood showed that monoclonal antibodies to HNL only react with neutrophils and not with other types of leukocytes. Immunocytochemistry on plastic-embedded sections and on frozen sections of lung tissue showed that a cocktail of six monoclonal antibodies to HNL specifically reacts with neutrophils and not with epithelial cells or macrophages. By immunoelectron microscopical studies performed on healthy human neutrophils after low temperature embedding in Lowicryl K4M following aldehyde fixation and partial dehydration, it could be shown that HNL colocalized with lactoferrin (a known marker for secondary or specific granules) and that myeloperoxidase was localized in the primary or azurophil granules. The results confirm that HNL is a unique component of the secondary granules of the neutrophil granulocyte.
嗜中性粒细胞是一种具有细胞毒性且可能损伤组织的细胞,参与多种炎症性疾病中所见的破坏过程和症状。已引入灵敏的免疫测定法来测量血液和其他体液中各种炎症细胞的特定分泌蛋白水平。目的是为每种细胞类型开发高度特异性的标志物。免疫测定法获得的结果表明,人嗜中性粒细胞脂质运载蛋白(HNL)是嗜中性粒细胞特有的一种蛋白质。本研究通过流式细胞术和免疫细胞化学研究了HNL作为外周血和肺组织中嗜中性粒细胞标志物的特异性。外周血的流式细胞术和免疫细胞化学显示,针对HNL的单克隆抗体仅与嗜中性粒细胞反应,而不与其他类型的白细胞反应。肺组织塑料包埋切片和冷冻切片的免疫细胞化学显示,六种针对HNL的单克隆抗体混合物特异性地与嗜中性粒细胞反应,而不与上皮细胞或巨噬细胞反应。通过对醛固定和部分脱水后低温包埋于Lowicryl K4M中的健康人嗜中性粒细胞进行免疫电子显微镜研究,可以表明HNL与乳铁蛋白(一种已知的次级或特异性颗粒标志物)共定位,并且髓过氧化物酶定位于初级或嗜天青颗粒中。结果证实HNL是嗜中性粒细胞次级颗粒的独特成分。