【摘要】 目的建立适用于临床的流式细胞术(FCM)检测抗中性粒细胞抗体(ANA)的方法,并作临床初步应用。方法采用FCM,以前向散射和侧向散射参数区分全血标本中的淋巴细胞和粒细胞,设定粒细胞门,用CD16b-藻红蛋白(PE)和CD177-别藻蓝蛋白(APC)单克隆抗体分别检测中性粒细胞中表面表达CD16b和CD177抗原细胞的百分率。用所建方法检测健康对照32名,确定参考范围,以此作为判断受检者血液中抗CD16b或CD177抗体存在与否的依据。检测外周血白细胞(WBC)总数<4.0×109/L且中性粒细胞绝对计数(ANC)<2.8×109/L的59例患者的CD16b和CD177百分率,并对其中中性粒细胞表达CD16b和/或CD177百分率低于参考范围(即判为相应抗体阳性)的患者给予激素治疗,观察疗效。结果健康人群中性粒细胞中表达CD16b和CD177的百分率分别为98.36%±1.53%和74.95%±11.07%,据此分别确定了健康人群中性粒细胞表达CD16b和CD177的参考范围分别为CD16b>95.36%、CD177>53.25%(即>x珋-1.96s)。59例患者中有23例(39.0%)ANA阳性,其中ANC<2.0×109/L患者的ANA阳性率(50.0%)远高于ANC>2.0×109/L患者(27.6%)。ANA阳性患者经激素治疗后,WBC和ANC上升。结论 FCM检测抗中性粒细胞CD16b和CD177抗体是自身免疫性中性粒细胞减少症(AIN)实验简便、快捷的诊断方法,有助于AIN的诊断、鉴别诊断和疗效判断。更多还原

【Abstract】 Objective To establish the flow cytometry method(FCM) for detecting anti-neutrophil antibody(ANA) and primarily apply it for testing ANA in patients with neutropenia.Methods Lymphocyte and granulocyte in whole blood were distinguished by forward scatter and side scatter parameters by FCM,and the granulocyte gate was set up.CD16b and CD177 antigens on neutrophil surface in whole blood samples were detected by CD16b-phycoerythrin(PE) and CD177-allophycocyanin(APC) monoclonal antibody respectively.The whole blood samples from 32 healthy subjects were detected by the established method to determine the reference ranges of CD16b and CD177,which was the basis to judge if the anti-CD16b or anti-CD177 antibodies existed in blood.The whole blood samples′ percentages of CD16b and CD177 from 59 patients with white blood cell(WBC) count <4.0×109/L and absolute neutrophil count(ANC) <2.8×109/L were detected by the method.The patients with CD16b and/or CD177 on neutrophil expression below the reference ranges(the corresponding antibody positive) were treated with hormone in order to the curative effect observation.Results The percentages of CD16b and CD177 expression on neutrophil in healthy controls were 98.36%±1.53% and 74.95±11.07%,and the reference ranges of the CD16b and CD177 expression on neutrophil were >95.36% and 53.25%(-1.96s),respectively.The 23(39.0%) of 59 patients were ANA positive,and the positive rate of ANA in the patients with ANC <2.0×109/L(50.0%) was higher than that in the patients with ANC >2.0×109/L(27.6%).The WBC and ANC of the patients with ANA positive were elevated after hormone therapy.Conclusions The FCM for detecting anti-neutrophil CD16b and CD177 antibodies is simple,fast and well applicable for the laboratory diagnosis and curative effect observation of autoimmune neutropenia(AIN).更多还原