Orfao A, Escribano L, Villarrubia J, Velasco J L, Cerveró C, Ciudad J, Navarro J L, San Miguel J F
Servicio Central de Citometria, Hospital Clínico Universitario, Salamanca, Spain.
Am J Pathol. 1996 Nov;149(5):1493-9.
In the present paper we have used a three-color immunofluorescence procedure combined with flow cytometry cell analysis and sorting for the identification and enumeration of human mast cells in both normal and pathological bone marrow samples. Our results show that bone marrow mast cells are clearly identifiable on the basis of their light-scatter properties and strong CD117 expression. These cells were negative for the CD34, CD38, and BB4 antigens. In addition, they were CD33+ and displayed a high reactivity for the anti-IgE monoclonal antibody. The identity of the CD117-strong+ cells (mast cells) was confirmed by both microscopic examination and flow cytometry analysis. The overall frequency of mast cells in the bone marrow samples analyzed in the present study was constantly lower than 1%. The lowest frequencies corresponded to normal human bone marrow samples (0.0080 +/- 0.0082%) and the highest to those patients suffering from indolent systemic mast cell disease (0.40 +/- 0.13%). In summary, our results show that the identification and enumeration of bone marrow mast cells can be achieved using multiparametric flow cytometry. Moreover, once identified, mast cells are suitable for being characterized from the phenotypic and the functional point of view, facilitating the comparison between normal and abnormal mast cells.
在本论文中，我们使用了三色免疫荧光程序结合流式细胞术细胞分析和分选技术，对正常和病理骨髓样本中的人肥大细胞进行鉴定和计数。我们的结果表明，骨髓肥大细胞可根据其光散射特性和强烈的CD117表达清晰地鉴定出来。这些细胞对CD34、CD38和BB4抗原呈阴性。此外，它们为CD33阳性，并对抗IgE单克隆抗体表现出高反应性。通过显微镜检查和流式细胞术分析均证实了CD117强阳性细胞（肥大细胞）的身份。在本研究中分析的骨髓样本中，肥大细胞的总体频率始终低于1%。最低频率对应于正常人类骨髓样本（0.0080±0.0082%），最高频率对应于患有惰性系统性肥大细胞病的患者（0.40±0.13%）。总之，我们的结果表明，使用多参数流式细胞术可以实现骨髓肥大细胞的鉴定和计数。此外，一旦鉴定出来，肥大细胞就适合从表型和功能角度进行表征，便于比较正常和异常肥大细胞。
