急性髓系白血病患者外周血粒细胞、单核细胞膜GPI锚链和红细胞表面CD59表达水平的研究
|
摘要
目的通过流式细胞术研究急性髓系白血病(AML)患者粒细胞、单核细胞糖基磷脂酰肌醇(glycosyl-phosphatidylinositol,GPI)锚链和红细胞CD59表达水平及其临床意义。方法该研究纳入了36例并发血管内溶血的AML患者及21例阵发性睡眠性血红蛋白尿(paroxysmal nocturnal hemoglobinuria,PNH)患者,依据国际临床流式细胞协会推荐的PNH流式诊断指南,分别对受试对象的红细胞和有核细胞进行PNH克隆检测,统计分析AML患者粒细胞、单核细胞及红细胞PNH克隆的水平。结果 AML组患者与PNH组患者相比较无明显红细胞CD59表达缺陷(I型红细胞98.49±4.61vs61.41±25.86,Ⅱ型红细胞1.33±4.55vs 16.82±19.92,Ⅲ型红细胞0.17±0.85vs 21.71±21.12,差异均有统计学意义(t=6.43,3.41,4.71,均P<0.001);粒细胞、单核细胞缺陷克隆检测存在但与PNH组比较则明显减少(粒细胞26.46±23.36vs 73.12±28.41,单核细胞35.80±27.80vs 78.37±25.03,差异有统计学意义(t=4.59,3.76,均P<0.001)。结论 AML患者有核细胞中可出现FLAER阴性的PNH克隆,但未见CD59表达缺陷的红细胞,可与PNH区分。
Objective To study the expression levels and the clinical significance of glycosyl-phosphatidyl inositol(GPI)and CD59 in AML patients by flow cytometry(FCM).Methods From 2014 to 2018,36 AML patients combined with intravascular hemolysis from Shanghai Ruijin Hospital were studied in this study,while 21 cases of paroxysmal nocturnal hemoglobinuria(PNH)patients were included as control.Multiparameter FCM was used to detect the expression levels of GPI anchor chain of Fluorescent-labeled aerophilic plasminogen(FLAER)treated granulocytes and monocytes in peripheral blood samples.FCM was performed to evaluate the expression level of CD59 of erythrocyte.Results Compared with PNH group patients,AML group patients had no obvious deficiency in CD59 expression(type I erythrocyte 98.49±4.61 vs 61.41±25.86,typeⅡerythrocyte 1.33±4.55 vs 16.82±19.92,typeⅢerythrocyte 0.17±0.85 vs 21.71±21.12),the differences were ststistically significant(t=6.43,3.41,4.71,all P<0.001).And the detection of granulocyte and mononuclear cell defects by cloning was found,but it was significantly reduced compared with PNH group patients(granulocyte 26.46±23.36 vs 73.12±28.41,monocyte 35.80±27.80 vs 78.37±25.03),the differences were statisyically significant(t=4.59,3.76,all P<0.001).Conclusion FLAER negative PNH clones could be found in nucleated cells of AML patients,while no CD59 expression defects could be found in erythrocytes,which could be differentiated from PNH.
Objective To study the expression levels and the clinical significance of glycosyl-phosphatidyl inositol(GPI)and CD59 in AML patients by flow cytometry(FCM).Methods From 2014 to 2018,36 AML patients combined with intravascular hemolysis from Shanghai Ruijin Hospital were studied in this study,while 21 cases of paroxysmal nocturnal hemoglobinuria(PNH)patients were included as control.Multiparameter FCM was used to detect the expression levels of GPI anchor chain of Fluorescent-labeled aerophilic plasminogen(FLAER)treated granulocytes and monocytes in peripheral blood samples.FCM was performed to evaluate the expression level of CD59 of erythrocyte.Results Compared with PNH group patients,AML group patients had no obvious deficiency in CD59 expression(type I erythrocyte 98.49±4.61 vs 61.41±25.86,typeⅡerythrocyte 1.33±4.55 vs 16.82±19.92,typeⅢerythrocyte 0.17±0.85 vs 21.71±21.12),the differences were ststistically significant(t=6.43,3.41,4.71,all P<0.001).And the detection of granulocyte and mononuclear cell defects by cloning was found,but it was significantly reduced compared with PNH group patients(granulocyte 26.46±23.36 vs 73.12±28.41,monocyte 35.80±27.80 vs 78.37±25.03),the differences were statisyically significant(t=4.59,3.76,all P<0.001).Conclusion FLAER negative PNH clones could be found in nucleated cells of AML patients,while no CD59 expression defects could be found in erythrocytes,which could be differentiated from PNH.
引文
[1]龚亚文,何广胜.阵发性睡眠性血红蛋白尿症的临床进展[J].中国实验血液学杂志,2013,21(6):1627-1630.GONG Yawen,HE Guangsheng.Clinical progres of paroxysmal nocturnal hemoglobinuria[J].Journal of Experimental Hematology,2013,21(6):1627-1630.
[2]梁悦怡,谢守军.FLAER多参数检测PNH克隆的意义[J].国际检验医学杂志,2016,37(8):1139-1141.LIANG Yueyi,XIE Shoujun.Significance of FLAERmulti-parameter detection of PNH cloning[J].International Journal of Laboratory Medicine,2016,37(8):1139-1141.
[3]刘淑媛,万腊根,闻芳,等.FLAER检测及其在阵发性睡眠性血红蛋白尿症诊断中的意义[J].实验与检验医学,2015(1):4-6,15.LIU Shuyuan,WAN Lagen,WEN Fang,et al.Significance of FLAER detection in the diagnosis of paroxysmal noctutnal hemoglobinuria[J].Experimental and Laboratory Medicine,2015(1):4-6,15.
[4]ALGHASHAM N,ABULKHAIR Y,KHALIL S.Flow cytometry screening for paroxysmal nocturnal hemoglobinuria:A single-center experience in Saudi A-rabia[J].Cytometry Part B Clinical Cytometry,2015,88(6):389-394.
[5]DAHMANI A,ROUDOT H,CYMBALISTA F,et al.Evaluation of fluorescently labeled aerolysin as a new kind of reagent for flow cytometry tests optimization of use of FLAER,hints,and limits[J].American Journal of Clinical Pathology,2016,145(3):407-417.
[6]韩秀蕊,杨娣娣,王九菊,等.136例急性白血病免疫表型特点及临床意义[J].现代检验医学杂志,2014,29(4):91-93.HAN Xiurui,YANG Didi,WANG Jiu-ju,et al.Immunophenotypie characteristics and clinical significance of 136patients with acute leukemia[J].Journal of Modern Laboratory Medicine,2014,29(4):91-93.
[7]袁莉,杨玉琮,陈葳.93例急性白血病流式细胞术免疫分型的特点分析[J].现代检验医学杂志,2014,29(2):127-129.YUAN Li,YANG Yuzong,CHEN Wei.Analysis of immunophenotypic features by flow cytometer in 93cases with acute leukemia[J].Journal of Modern Laboratory Medicine,2014,29(2):127-129.
[8]杨柯,郭晓宇,欧剑锋,等.外周血粒细胞CD55,CD59和FLAER检测在贫血及PNH诊断中的意义[J].现代检验医学杂志,2017,32(3):6-10.YANG Ke,GUO Xiaoyu,OU Jianfeng,et al.Diagnostic significance of detecting peripheral blood granulocyte CD55,CD59and FLAER in anemia and PNH[J].Journal of Modern Laboratory Medicine,2017,32(3):6-10.
[9]谢亚荣,任方刚,张娜,等.CD55CD59检测在阵发性睡眠性血红蛋白尿诊断中的意义[J].中国药物与临床,2016,16(5):756-758.XIE Yarong,REN Fanggang,ZHANG Na,et al.Significance of CD55CD59detection in the diagnosis of paroxysmal sleep hemoglobinuria[J].Chinese Remedies and Clinical,2016,16(5):756-758.
[10]赵阿兰,陈智超,张青艳,等.有核细胞嗜水气单胞菌溶素变异体流式法:一种更敏感稳定的阵发性睡眠性血红蛋白尿症检测方法[J].临床内科杂志,2013,30(3):199-202.ZHAO Alan,CHEN Zhichao,ZHANG Qing-yan,et al.Karyocyte base FLAER flow cytometry:a more stable and sensitive means to detect PNH[J].Journal of Clinical Internal Medicine,2013,30(3):199-202.
[2]梁悦怡,谢守军.FLAER多参数检测PNH克隆的意义[J].国际检验医学杂志,2016,37(8):1139-1141.LIANG Yueyi,XIE Shoujun.Significance of FLAERmulti-parameter detection of PNH cloning[J].International Journal of Laboratory Medicine,2016,37(8):1139-1141.
[3]刘淑媛,万腊根,闻芳,等.FLAER检测及其在阵发性睡眠性血红蛋白尿症诊断中的意义[J].实验与检验医学,2015(1):4-6,15.LIU Shuyuan,WAN Lagen,WEN Fang,et al.Significance of FLAER detection in the diagnosis of paroxysmal noctutnal hemoglobinuria[J].Experimental and Laboratory Medicine,2015(1):4-6,15.
[4]ALGHASHAM N,ABULKHAIR Y,KHALIL S.Flow cytometry screening for paroxysmal nocturnal hemoglobinuria:A single-center experience in Saudi A-rabia[J].Cytometry Part B Clinical Cytometry,2015,88(6):389-394.
[5]DAHMANI A,ROUDOT H,CYMBALISTA F,et al.Evaluation of fluorescently labeled aerolysin as a new kind of reagent for flow cytometry tests optimization of use of FLAER,hints,and limits[J].American Journal of Clinical Pathology,2016,145(3):407-417.
[6]韩秀蕊,杨娣娣,王九菊,等.136例急性白血病免疫表型特点及临床意义[J].现代检验医学杂志,2014,29(4):91-93.HAN Xiurui,YANG Didi,WANG Jiu-ju,et al.Immunophenotypie characteristics and clinical significance of 136patients with acute leukemia[J].Journal of Modern Laboratory Medicine,2014,29(4):91-93.
[7]袁莉,杨玉琮,陈葳.93例急性白血病流式细胞术免疫分型的特点分析[J].现代检验医学杂志,2014,29(2):127-129.YUAN Li,YANG Yuzong,CHEN Wei.Analysis of immunophenotypic features by flow cytometer in 93cases with acute leukemia[J].Journal of Modern Laboratory Medicine,2014,29(2):127-129.
[8]杨柯,郭晓宇,欧剑锋,等.外周血粒细胞CD55,CD59和FLAER检测在贫血及PNH诊断中的意义[J].现代检验医学杂志,2017,32(3):6-10.YANG Ke,GUO Xiaoyu,OU Jianfeng,et al.Diagnostic significance of detecting peripheral blood granulocyte CD55,CD59and FLAER in anemia and PNH[J].Journal of Modern Laboratory Medicine,2017,32(3):6-10.
[9]谢亚荣,任方刚,张娜,等.CD55CD59检测在阵发性睡眠性血红蛋白尿诊断中的意义[J].中国药物与临床,2016,16(5):756-758.XIE Yarong,REN Fanggang,ZHANG Na,et al.Significance of CD55CD59detection in the diagnosis of paroxysmal sleep hemoglobinuria[J].Chinese Remedies and Clinical,2016,16(5):756-758.
[10]赵阿兰,陈智超,张青艳,等.有核细胞嗜水气单胞菌溶素变异体流式法:一种更敏感稳定的阵发性睡眠性血红蛋白尿症检测方法[J].临床内科杂志,2013,30(3):199-202.ZHAO Alan,CHEN Zhichao,ZHANG Qing-yan,et al.Karyocyte base FLAER flow cytometry:a more stable and sensitive means to detect PNH[J].Journal of Clinical Internal Medicine,2013,30(3):199-202.