van der Schoot C E, Huizinga T W, van 't Veer-Korthof E T, Wijmans R, Pinkster J, von dem Borne A E
Department of Immunological Hematology, University of Amsterdam, The Netherlands.
Blood. 1990 Nov 1;76(9):1853-9.
Paroxysmal nocturnal hemoglobinuria (PNH) is a disease that affects not only red cells, but other blood cells as well. The common defect is supposed to be an acquired deficiency of glycosyl-phosphatidylinositol (GPI)-anchored membrane proteins, which may be present already at the hematopoietic stem cell level. Recently, a panel of monoclonal antibodies (MoAbs) has become available directed against various GPI-linked membrane proteins. This makes it possible to study various cell lineages for the deficiency of such proteins in PNH in more detail. Using cytofluorography, we could show that the granulocytes of 20 different PNH patients miss not only GPI-linked FcRIII (CD16 antigen), but also three other GPI-linked proteins, ie, CD24 antigen, CD67 antigen and a granulocyte-specific 50 to 80 Kd antigen. The affected granulocytes were not only neutrophils but also eosinophils, as was found in a more detailed analysis of three patients. Moreover, in all 10 PNH patients tested, the monocytes were found to be deficient for the GPI-linked CD14 antigen, and we found with CD24 and CD55 (DAF) antibodies that lymphocytes may be involved as well. However, abnormal B and T lymphocytes were detected only in a subset of patients (2 of 10 tested). The uniform deficiency of GPI-linked proteins of granulocytes allows the introduction of a new diagnostic cytofluorometric assay for PNH with MoAbs against GPI-linked granulocytic antigens. This test was positive in all PNH patients studied and not in a group of 40 control patients or 50 normal donors, with the exception of three of 16 aplastic anemia (AA) patients. In the three AA patients, subpopulations (10% to 20%) of PNH granulocytes could be detected, whereas these patients had a negative acidified serum (Ham) test. This indicates that the new test is more sensitive than the Ham test and allows the early diagnosis of PNH in AA. An advantage of the neutrophil assay is that, in contrast to the Ham test, it is not influenced by recent red-cell transfusions. Moreover, it is possible to quantify the number of affected cells by single cell analysis.
阵发性睡眠性血红蛋白尿（PNH）是一种不仅影响红细胞，还影响其他血细胞的疾病。常见缺陷被认为是糖基磷脂酰肌醇（GPI）锚定膜蛋白的后天缺乏，这种缺乏可能在造血干细胞水平就已存在。最近，一组针对各种GPI连接膜蛋白的单克隆抗体（MoAbs）已可获得。这使得更详细地研究PNH中各种细胞谱系中此类蛋白的缺乏情况成为可能。通过细胞荧光术，我们能够表明20例不同PNH患者的粒细胞不仅缺失GPI连接的FcRIII（CD16抗原），还缺失其他三种GPI连接蛋白，即CD24抗原、CD67抗原和一种粒细胞特异性的50至80 Kd抗原。在对三名患者进行更详细分析时发现，受影响的粒细胞不仅有中性粒细胞，还有嗜酸性粒细胞。此外，在所有10例接受检测的PNH患者中，发现单核细胞缺乏GPI连接的CD14抗原，并且我们用CD24和CD55（衰变加速因子，DAF）抗体发现淋巴细胞也可能受累。然而，仅在一部分患者（10例检测患者中的2例）中检测到异常的B和T淋巴细胞。粒细胞GPI连接蛋白的一致缺乏使得可以引入一种新的针对PNH的诊断性细胞荧光测定法，使用针对GPI连接粒细胞抗原的MoAbs。该检测在所有研究的PNH患者中呈阳性，而在40例对照患者或50例正常供者中均为阴性，但16例再生障碍性贫血（AA）患者中有3例除外。在这3例AA患者中，可检测到PNH粒细胞亚群（10%至20%），而这些患者的酸化血清（Ham）试验为阴性。这表明新检测比Ham试验更敏感，并且能够在AA中早期诊断PNH。中性粒细胞检测的一个优点是，与Ham试验不同，它不受近期红细胞输血的影响。此外，通过单细胞分析可以量化受影响细胞的数量。
