Sakamoto N, Sakamoto H, Tanaka S, Oryu M, Ogawa Y
2nd Department of Pathology, Kagawa Medical University, Kagawa Pref., Japan.
J Leukoc Biol. 1998 Nov;64(5):631-5. doi: 10.1002/jlb.64.5.631.
Large and small macromolecular activators of phagocytosis from platelets (I-MAPP and s-MAPP, respectively), which function via the neutrophilic Fc gamma receptors (Fc gammaR) were refined from platelet release products by gel filtration and affinity chromatography with the use of an anti transferrin antibody column and the mechanism of phagocytosis activation was investigated. Flow cytometry revealed that 1-MAPP and s-MAPP did not increase the expression of neutrophilic Fc gammaRII (CD32) and Fc gammaRIII (CD16) antigens, whereas rosette formation of neutrophils with rabbit IgG-sensitized sheep erythrocytes (EA) in the presence of anti Fc gammaR antibodies suggested that both MAPPs increase the binding ability of Fc gammaRII. On the other hand, the enhancing effect of I-MAPP and s-MAPP on neutrophilic phagocytosis disappeared with the increase in phagocytosis by the phosphate-buffered saline control neutrophils when they were centrifuged with EA before incubation for phagocytosis. The enhanced phagocytosis, both by the two MAPPs and centrifugation, was canceled by treatment of the neutrophils with anti-CD32 Fab. The phagocytosis activatory effects of MAPP on neutrophils were canceled by anti-CD71 monoclonal antibody but not by transferrin.
分别通过凝胶过滤和亲和色谱法,利用抗转铁蛋白抗体柱从血小板释放产物中提纯了通过嗜中性粒细胞Fcγ受体(FcγR)发挥作用的血小板来源的大小两种吞噬作用大分子激活剂(分别为I-MAPP和s-MAPP),并对吞噬作用的激活机制进行了研究。流式细胞术显示,I-MAPP和s-MAPP并未增加嗜中性粒细胞FcγRII(CD32)和FcγRIII(CD16)抗原的表达,而在抗FcγR抗体存在的情况下,嗜中性粒细胞与兔IgG致敏绵羊红细胞(EA)形成的玫瑰花结表明,两种MAPP均增加了FcγRII的结合能力。另一方面,当用EA进行离心处理后再进行吞噬作用孵育时,随着磷酸盐缓冲盐水对照嗜中性粒细胞吞噬作用的增加,I-MAPP和s-MAPP对嗜中性粒细胞吞噬作用的增强作用消失。两种MAPP和离心处理所增强的吞噬作用,均会被用抗CD32 Fab处理嗜中性粒细胞所消除。MAPP对嗜中性粒细胞的吞噬作用激活效应会被抗CD71单克隆抗体消除,但不会被转铁蛋白消除。