Mutasa H C
Department of Haematology, Cambridge University Clinical School, United Kingdom.
Cell Tissue Res. 1989 Oct;258(1):111-7. doi: 10.1007/BF00223150.
In this study immuno-electron microscopy was used to assay, semi-quantitatively, the granule contents of elastase, lactoferrin, lysozyme and myeloperoxidase in human peripheral blood neutrophils from 13 chronic myeloid leukaemia patients in the chronic phase of the disease and from normal non-smoking donors. The fixation conditions that adequately preserved the antibody binding capacities of these antigens and reasonably preserved the ultrastructure of the neutrophils were selected by light-microscopic immunoperoxidase cytochemistry on cytospin smears. Immunogold cytochemistry on LR White resin sections localised elastase and myeloperoxidase to the primary granules, lactoferrin to the secondary granules and lysozyme to both types of granule. When applicable, peroxidase cytochemistry was combined with immunogold staining making it easier to distinguish the primary from the secondary granules. A comparison of the immunolabelling density values obtained for the leukaemic and normal states revealed no significant abnormalities in the immunoreactivity patterns for any of these neutrophil granule antigens in the leukaemic patients. All 13 patients gave normal immunostaining reactivities for these neutrophil granule proteins. Consequently the distribution patterns of these proteins, as shown in this study, cannot be used as indices in distinguishing chronic myeloid leukaemic neutrophils from normal neutrophils.
在本研究中，采用免疫电子显微镜对13例慢性粒细胞白血病慢性期患者及正常不吸烟供者的人外周血中性粒细胞中的弹性蛋白酶、乳铁蛋白、溶菌酶和髓过氧化物酶的颗粒内容物进行了半定量分析。通过对细胞涂片进行光镜免疫过氧化物酶细胞化学，选择了能充分保留这些抗原的抗体结合能力并合理保留中性粒细胞超微结构的固定条件。对LR White树脂切片进行免疫金细胞化学分析，结果显示弹性蛋白酶和髓过氧化物酶定位于初级颗粒，乳铁蛋白定位于次级颗粒，溶菌酶定位于两种颗粒。在适用的情况下，将过氧化物酶细胞化学与免疫金染色相结合，使得区分初级颗粒和次级颗粒更加容易。对白血病状态和正常状态下获得的免疫标记密度值进行比较，结果显示白血病患者中这些中性粒细胞颗粒抗原的免疫反应模式均无明显异常。所有13例患者对这些中性粒细胞颗粒蛋白的免疫染色反应均正常。因此，本研究中所示的这些蛋白质的分布模式不能用作区分慢性粒细胞白血病中性粒细胞和正常中性粒细胞的指标。
