Diagnosis and carrier detection of chronic granulomatous disease in five families by flow cytometry
Diagnosis and carrier detection of chronic granulomatous disease in five families by flow cytometry
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DOI：
10.1159/000237660
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发表时间：
1997-10-01
影响因子：
2.8
通讯作者：
Turner, CP
中科院分区：
文献类型：
--
作者：
Crockard, AD;Thompson, JM;Turner, CP
背景：应用流式细胞术测定吞噬细胞呼吸爆发(ROB)活性来研究慢性肉芽肿病(CGD)，可能会改进对患者和携带者的实验室检测，并表明分子缺陷的性质。为评价流式细胞术对5个CGD家系的诊断能力，对5个CGD家系(2个为X-CGD，3个为常染色体隐性CGD)的中性粒细胞经佛波酯(PMA)诱导的中性粒细胞ROB进行检测。结果：W-CGD患者中性粒细胞无反应性。女性携带者表现出高强度和低强度的双荧光峰，分别代表正常人群和异常人群。1例X-CGD经骨髓移植成功治疗的男孩，ROB活性正常。3例常染色体隐性遗传性CGD患者的ROB活性与其双亲和同胞相比明显降低，其流式细胞仪检测结果与不同的分子缺陷相关。缺乏NADPH氧化酶复合体的p22(Phox)膜组分导致呼吸爆发活性显着降低，这与在X-CGD中观察到的水平相当。然而，在p22(Phox)水平降低的患者和胞浆P47(Phox)成分缺乏的患者中，观察到呼吸爆发活动水平降低但可检测到的水平。结论：DHR流式细胞术为CGD提供了一种敏感的诊断筛选试验，并可能进一步提供可能的潜在分子缺陷的指示。
Background: The application of flow cytometric assays, for determination of phagocyte respiratory burst (ROB) activity, to the investigation of chronic granulomatous disease (CGD) may lead to improved laboratory detection of patients and carriers and indicate the nature of the molecular defect. To evaluate the diagnostic capability of flow cytometry an investigation of 5 CGD families was undertaken.Methods: Phorbol myristate acetate (PMA)-induced neutrophil ROB was determined using dihydrorhodamine 123 (DHR) and flow cytometric analysis in 26 members of 5 CGD families (2:X-CGD;3:autosomal recessive CGD).Results: Neutrophils from W-CGD patients displayed absence of reactivity. Female carriers demonstrated dual fluorescence peaks of high and low intensity indicative of normal and abnormal populations, respectively. Normal ROB activity was observed in a boy whose X-CGD was successfully treated by bone marrow transplantation. Reduced ROB activity was observed in 3 patients with autosomal-recessive CGD compared with their parents and siblings, The patterns of flow cytometric reactivity correlated with the different molecular defects identified. Absence of the p22(phox) membrane component of the NADPH oxidase complex resulted in a significantly reduced level of respiratory burst activity which was comparable to that observed in X-CGD. whereas reduced but detectable levels of respiratory burst activity were observed in a patient with diminished levels of p22(phox) and in a patient with deficiency of the cytosolic p47(phox) component.Conclusions: The DHR flow cytometric assay offers a sensitive diagnostic screening test for CGD and furthermore may provide an indication of the likely underlying molecular defect.