Lanza F, Latorraca A, Musto P, Ferrari L, Moretti S, Zabucchi G, Carotenuto M, Castoldi G L
Institute of Hematology, University of Ferrara, Italy.
Ann Hematol. 1991 Aug;63(2):94-100. doi: 10.1007/BF01707280.
Neutrophil granulocytes from 12 subjects with primary myeloperoxidase (MPO) deficiency (six totally deficient) and 16 patients with secondary partial MPO deficiency were tested using two different anti-MPO antibodies, in combination with either a flow-cytometric technique or an immunoalkaline phosphatase staining method. Results demonstrated three different cytofluorimetric patterns of immunoreactivity with the MPO protein: (a) a bright MPO antigenic expression, typical of patients with secondary MPO deficiency (comparable to that observed in the control group); (b) a medium MPO antigenic expression, typical of subjects with primary partial MPO deficiency; and (c) a dim MPO antigenic expression, characteristic of individuals with hereditary total MPO deficiency. No significant differences in granulocyte MPO reactivity were demonstrated for the two antibodies. Furthermore, in two individuals with complete primary enzyme deficiency, the single histogram analysis of MPO fluorescence determined by flow cytometry seemed to show that only 38% (case 1) and 44% (case 2) of neutrophils were reactive with the anti-MPO antibodies: the use of multiple histogram analysis in combination with Kolmogorov-Smirnov statistics allowed us to demonstrated that all the cells express a low density of MPO antigen. These data were more or less confirmed by the APAAP labeling method, which showed a reduced straining only in subjects with primary deficiency, while all patients with secondary deficiency had scores similar to those observed in controls (healthy subjects). Compared with the immunoenzymatic technique, the flow-cytometric procedure showed a higher sensitivity to MPO, being able to estimate even minor decreases in neutrophil MPO antigenic expression, as previously postulated by other authors. This work suggests that patients with primary MPO deficiency have different amounts of MPO antigens in the neutrophil granulocytes, and the levels of MPO fluorescence seem to decline concurrently with the enzyme activity, thereby suggesting the presence of a diminished MPO production. In contrast, the normal antigenic reactivity of neutrophils from patients with acquired MPO deficiency indicates the presence of a functionally inactive form of the enzyme.
使用两种不同的抗髓过氧化物酶（MPO）抗体，并结合流式细胞术或免疫碱性磷酸酶染色方法，对12名原发性髓过氧化物酶（MPO）缺乏症患者（6名完全缺乏）的中性粒细胞和16名继发性部分MPO缺乏症患者进行了检测。结果显示了与MPO蛋白免疫反应性的三种不同细胞荧光模式：（a）明亮的MPO抗原表达，是继发性MPO缺乏症患者的典型表现（与对照组观察到的相似）；（b）中等MPO抗原表达，是原发性部分MPO缺乏症患者的典型表现；（c）暗淡的MPO抗原表达，是遗传性完全MPO缺乏症个体的特征。两种抗体在粒细胞MPO反应性方面未显示出显著差异。此外，在两名原发性酶完全缺乏的个体中，通过流式细胞术对MPO荧光进行的单直方图分析似乎显示，只有38%（病例1）和44%（病例2）的中性粒细胞与抗MPO抗体有反应：使用多重直方图分析并结合Kolmogorov-Smirnov统计方法使我们能够证明所有细胞均表达低密度的MPO抗原。这些数据或多或少得到了碱性磷酸酶抗碱性磷酸酶（APAAP）标记法的证实，该方法显示仅原发性缺乏症患者的染色减少，而所有继发性缺乏症患者的得分与对照组（健康受试者）观察到的相似。与免疫酶技术相比，流式细胞术对MPO的敏感性更高，能够像其他作者先前推测的那样，估计中性粒细胞MPO抗原表达的微小下降。这项研究表明，原发性MPO缺乏症患者的中性粒细胞中MPO抗原的含量不同，MPO荧光水平似乎与酶活性同时下降，从而表明存在MPO产生减少的情况。相比之下，获得性MPO缺乏症患者中性粒细胞的正常抗原反应性表明存在该酶的功能无活性形式。
