Cui Wei, Lin Qisui, Zhang Zhinan
Department of Laboratory Medicine, Peking Union Medical College Hospital, Beijing 100730, China.
Chin Med J (Engl). 2002 Feb;115(2):206-8.
To determine whether affected reticulocytes could be a reliable marker for the diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), we analyzed CD59-antigen expression on the membranes of reticulocytes and erythrocytes.
We studied 10 PNH patients and 5 healthy volunteers by two-color flow cytometry with a membrane permeable fluorescent dye, thiazole orange (TO), and anti-CD59 monoclonal antibodies (MoAb). TO was introduced to gate reticulocytes and anti-CD59 MoAb were used to identify glycosylphosphatidylinositol (GPI)-deficient cells.
Cells from healthy individuals were only CD59 positive. However, in all PNH patients, CD59-antigen expression could be divided into 3 types: type I cells (CD59 normally positive), type II cells (CD59 partly positive) and type III cells (CD59 negative). The majority of reticulocytes belonged to type III cells, GPI-deficient cells (61.0%). In addition, the percentage of affected reticulocytes was higher than that of erythrocytes.
Analyzing PNH reticulocytes was important, because most patients had elevated numbers of reticulocytes, which represent more closely the recent erythroid output of BM. However, circulating mature erythrocytes were subject to complement-mediated intravascular lysis. Therefore, the percentage of abnormal erythrocytes may not accurately reflect the proliferation rate of normal and abnormal erythroid progenitor cells. Thus, affected reticulocytes could be a more reliable indicator for the diagnosis of PNH than mature erythrocytes.
为了确定受影响的网织红细胞是否可作为阵发性夜间血红蛋白尿（PNH）诊断的可靠标志物，我们分析了网织红细胞和红细胞膜上CD59抗原的表达。
我们使用膜通透性荧光染料噻唑橙（TO）和抗CD59单克隆抗体（MoAb），通过双色流式细胞术研究了10例PNH患者和5名健康志愿者。TO用于圈定网织红细胞，抗CD59 MoAb用于识别糖基磷脂酰肌醇（GPI）缺陷细胞。
健康个体的细胞仅CD59呈阳性。然而，在所有PNH患者中，CD59抗原表达可分为3种类型：I型细胞（CD59正常阳性）、II型细胞（CD59部分阳性）和III型细胞（CD59阴性）。大多数网织红细胞属于III型细胞，即GPI缺陷细胞（61.0%）。此外，受影响的网织红细胞百分比高于红细胞。
分析PNH网织红细胞很重要，因为大多数患者的网织红细胞数量增加，其更能代表近期骨髓的红系输出。然而，循环中的成熟红细胞会发生补体介导的血管内溶血。因此，异常红细胞的百分比可能无法准确反映正常和异常红系祖细胞的增殖率。因此，与成熟红细胞相比，受影响的网织红细胞可能是PNH诊断更可靠的指标。
