A monoclonal antibody to a human neutrophil-specific plasma membrane antigen. Effect of the antibody on the C3bi-mediated adherence by neutrophils and expression of the antigen during myelopoiesis.
A monoclonal antibody to a human neutrophil-specific plasma membrane antigen. Effect of the antibody on the C3bi-mediated adherence by neutrophils and expression of the antigen during myelopoiesis.
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DOI：
10.1084/jem.167.2.421
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发表时间：
1988-02-01
期刊：
影响因子：
--
通讯作者：
Michl J
中科院分区：
文献类型：
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作者：
Pytowski B;Easton TG;Valinsky JE;Calderon T;Sun T;Christman JK;Wright SD;Michl J
我们用环磷酰胺治疗对人淋巴细胞抗原有选择性耐受的小鼠，培养了一种单抗BH2-C6，它能与人中性粒细胞特异性的质膜抗原反应。间接免疫荧光显微镜、流式细胞术分离的荧光阳性和阴性细胞群的细胞化学分析，以及补体介导的mAb bh2 - c6处理的中性粒细胞的选择性杀伤，证明了这种特异性。免疫电镜显示了mAb BH2-C6对中性粒细胞特异性的其他证据，这表明与人类嗜酸性粒细胞缺乏反应性。用125i标记的BH2-C6对多形核白细胞的SDS-PAGE分离蛋白进行免疫印迹，鉴定出平均分子质量为157 kD的蛋白。结合研究表明，在饱和状态下，中性粒细胞在每个细胞中结合214,000个125I-BH2-C6分子。向中性粒细胞中添加mAb BH2-C6可显著减少与这些细胞结合的C3bi-opsonized绵羊红细胞（EIgMC3bi）的数量。大豆胰蛋白酶抑制剂（SBTI）的存在部分逆转了这种减少，表明这种抑制至少有一部分是由于BH2-C6刺激分泌的丝氨酸蛋白酶可能影响配体结合。用细胞荧光法对正常人骨髓细胞进行细胞化学分析，发现早幼粒细胞是首先在质膜上表达BH2-Ag的前体细胞。利用白血病细胞系HL-60，我们证明了只有粒细胞分化诱导剂、顺式维甲酸和二甲基氯唑烷能刺激BH2-Ag的表达。这些结果表明，BH2- Ag在骨髓单核细胞分化过程中的表达是嗜中性细胞谱系所独有的特性。
We have used mice selectively tolerized to antigens of human lymphocytes by treatment with cyclophosphamide to raise an mAb, BH2-C6, that reacts with a plasma membrane antigen specific for human neutrophils. This specificity is demonstrated by indirect immunofluorescence microscopy, cytochemical analysis of fluorescence- positive and -negative cell populations separated by flow cytometry, and by the selective, complement-mediated killing of mAb BH2-C6-treated neutrophils. Additional evidence for the neutrophil specificity of mAb BH2-C6 is shown by immunoelectron microscopy, which demonstrates a lack of reactivity with human eosinophils. Immunoblotting of SDS-PAGE- separated proteins of polymorphonuclear leukocytes with 125I-labeled BH2-C6 identifies protein with an average molecular mass of 157 kD. Binding studies show that, at saturation, neutrophils bind 214,000 molecules of 125I-BH2-C6 per cell. Addition of mAb BH2-C6 to neutrophils significantly reduces the number of C3bi-opsonized sheep erythrocytes (EIgMC3bi) bound by these cells. This reduction is partly reversed by the presence of soybean trypsin inhibitor (SBTI), indicating that at least one part of this inhibition is due to BH2-C6- stimulated secretion of a serine protease that may affect ligand binding. Cytochemical analysis of normal human bone marrow cells sorted by cytofluorimetry identifies the promyelocyte as the precursor cell that first expresses BH2-Ag on the plasma membrane. Using the leukemic cell line HL-60, we demonstrate that only inducers of granulocytic differentiation, cis-retinoic acid, and dimethyloxazolidine stimulate the expression of BH2-Ag. These results show that the expression of BH2- Ag during myelomonocytic differentiation is a property uniquely possessed by cells committed to the neutrophilic lineage.