人中性粒细胞防御素单克隆抗体制备及初步应用

Preparation and preliminary trial of monoclonal antibodies to human neutrophil defensins

目的 制备人中性粒细胞防御素 1单克隆抗体 (HNP 1McAb) ,并用HNP 1McAb建立直接免疫荧光法对正常人及一系列髓性白血病患者中性粒细胞进行免疫细胞化学染色。方法 用连续酸 尿素 聚丙烯酰胺凝胶电泳分离纯化人中性粒细胞防御素组份 (HNP 1～ 3) ,以纯化的HNP 1为抗原制备特异性的HNP 1McAb ,并建立了HNP 1McAb直接免疫荧光染色法。将分离纯化的正常人和兔外周血中性粒细胞、淋巴细胞 ,粒细胞白血病、淋巴细胞白血病患者的骨髓和外周血分别制成涂片 ,进行免疫荧光染色。结果 HNP 1McAb与正常人外周血中性粒细胞及急、慢性粒细胞白血病骨髓和外周血涂片的原、早幼、中幼、晚幼和成熟的中性粒细胞均呈特异的阳性反应。人红细胞、淋巴细胞及其他非粒细胞系统的有核细胞均呈阴性反应。对兔的中性粒细胞无交叉反应。结论 HNP 1可作为粒细胞系统的特异性分化标志 ,其相应的HNP

Objective To prepare the monoclonal antibodies to human neutrophil defensins-1(HNP-1 McAb) and develop a direct immunofluorescence method for immunocytochemical staining of neutrophils from a series of normal or myelogenous leukemia patients.Methods The components of neutrophil defesins HNP-1, 2,3 were isolated and purified by continuous acid-urea-polyacrilamide gel electrophoresis.The monoclonal antibodies (McAbs) specific against HNP-1 were prepared by using purified HNP-1 as antigen.A direct immunofluorescence method was developed by using the McAbs for immunofluorescent staining of the neutrophils in isolated normal human or rabbit peripheral blood smears and the neutrophils in both peripheral blood and bone marrow smears of granulocytic or lymphocytic leukemia patients.Results Specific positive reactions of HNP-1 were observed in the neutrophils of peripheral blood from normal human subjects and myeloblast,neutrophillic promyelocyte,myelocyte metamyelocyte and mature neutrophils of both peripheral blood and bone marrow from the patients with acute or chronic granulocytic leukemia.Negative results were obtained in human erythrocytes,lymphocytes and other non-granulocytic karyocytes, and also in rabbit neutrophils.Conclusions HNP-1 may be a specific differentiation marker of granulocytes and its monoclonal antibodies may be used for differential diagnosis and typing of myelogenous leukemia.