摘要
目的探讨阵发性睡眠性血红蛋白尿症（ＰＮＨ）患者网织红细胞膜上反应性溶血膜抑制物（ＣＤ５９）的表达对诊断ＰＮＨ的意义。方法对１０例ＰＮＨ患者及５例健康成人，采用流式细胞术间接免疫荧光双标记法检测外周血红系ＣＤ５９的表达，荧光染料噻唑橙用于测定网织红细胞，抗ＣＤ５９单抗检测糖化肌醇磷脂结合蛋白（ＧＰＩ锚蛋白）缺失的细胞。结果健康成人网织红细胞及红细胞膜上ＣＤ５９表达完全阳性，而ＰＮＨ患者ＣＤ５９表达分为三型，Ⅰ型细胞的ＣＤ５９完全阳性，Ⅱ型细胞ＣＤ５９部分阳性，Ⅲ型细胞ＣＤ５９完全阴性。１０例ＰＮＨ患者５例表现为两群即Ⅰ型细胞＋Ⅲ型细胞或Ⅱ型细胞＋Ⅲ型细胞，４例表现为三群即Ⅰ型细胞＋Ⅱ型细胞＋Ⅲ型细胞，１例红细胞表现为Ⅰ型细胞，而其网织红细胞为Ⅰ型细胞＋Ⅲ型细胞。绝大多数ＰＮＨ患者网织红细胞中Ⅲ型细胞占主要成分（６１．０％），同一患者病态网织红细胞高于病态成熟红细胞，ＲＥＴ／ＲＢＣⅢ或ＲＥＴ／ＲＢＣⅡ＋Ⅲ。
Objective Paroxysmal nocturnal hemoglobinuria (PNH) blood cells lack glycosylphosphatidylinositol(GPI) anchored membrane proteins, such as CD 59 . To indicate that affected reticulocytes could be a reliable marker for the diagnosis of PNH and for the evalation of erythropoiesis by PNH stem cell, we analyzed CD 59 antigen expression on the surface membranes of reticulocytes and erythrocytes.Method We studied 10 PNH patients and 5 healthy volunteers by two color flow cytometry with a membrane permeable fluorescent dye, thiazole orange (TO), to identify reticulocytes and CD 59 monoclonal antibodies to identify GPI deficient cells. Results Healthy individuals had no affected cells, which have only CD 59 normally positive patients. However, in all PNH CD 59 antigen expression was divided into three types: I type cell(CD 59 normally positive), Ⅱtype cell(CD 59 partly positive), and Ⅲ type cell (CD 59 negative). The majority of reticulocytes was completely GPI deficient, ie: Ⅲ type cells (61.0%). In addition, the number of affected reticulocytes was higher than affected erythrocytes in the same patients, ie, RET/RBC Ⅲ or RET/RBC Ⅱ+Ⅲ >1.Conclusion Analysis of PNH reticulocytes is important, because most patients have increased number of reticulocytes that represent the recent erythroid output of BM. Because circulating mature erythrocytes are related to complement mediated intravascular lysis, the percentage of abnormal erythrocytes may not accurately reflect the proliferative rates of normal and abnormal erythroid progenitor cells. Hence, affected reticulocytes could be a more reliable marker for the diagnosis of PNH than mature erythrocytes.
