CHARACTERIZATION OF THE CYTOLYTIC TRIGGER MOLECULES G7/PNK-E AS A MOLECULAR-COMPLEX ON THE SURFACE OF PORCINE PHAGOCYTES
CHARACTERIZATION OF THE CYTOLYTIC TRIGGER MOLECULES G7/PNK-E AS A MOLECULAR-COMPLEX ON THE SURFACE OF PORCINE PHAGOCYTES
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DOI：
10.1006/cimm.1995.1036
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发表时间：
1995-04-01
影响因子：
4.3
通讯作者：
KIM, YB
中科院分区：
文献类型：
--
作者：
ALLER, SC;CHO, DH;KIM, YB
G7和PNK-E单抗识别不同的猪NK细胞和粒细胞功能相关分子，通过重定向的细胞毒机制增强和诱导针对肿瘤细胞靶点的显著细胞杀伤反应。本研究表明，G7和PNK-E分子存在于猪中性粒细胞、单核细胞和肺泡巨噬细胞表面，是一种物理和功能相关的细胞溶解触发分子复合体。双色流式细胞术分析表明，大多数(如果不是全部)中性粒细胞G7和PNK-E抗原阳性，而单核细胞和PAM中既有G7和PNK-E阳性，也有G7阳性和PNK-E阴性亚群。单抗结合竞争实验表明，G7单抗可阻断PNK-E单抗与吞噬细胞的后续结合，提示PNK-E单抗与吞噬细胞表面存在物理结合。利用G7和PNK-E单抗进行的荧光共封盖实验清楚地证明了存在于猪中性粒细胞表面的G7和PNK-E抗原之间的物理联系。用G7和PNK-E单抗的F(ab‘)(2)片段预处理吞噬细胞表明，F(ab’)(2)G7单抗可阻断整个PNK-E单抗随后诱导的吞噬细胞介导的肿瘤细胞杀伤作用，但F(ab‘)(2)PNK-E不能阻断整个G7单抗随后诱导的吞噬细胞介导的肿瘤细胞杀伤作用。中性粒细胞和单核巨噬细胞用F(ab‘)(2)单抗片段预处理能阻断PNK-E单抗依赖的吞噬细胞内氧化猝发反应的后续激活，但用F(ab’)(2)PNK-E不能阻止吞噬细胞内氧化猝发反应的后续依赖于G7单抗的激活，这些数据加强了G7和PNK-E分子之间的物理和功能联系，最近发现G7抗原是Fc-Gamma RIII的猪同源基因，G7和PNK-E单抗识别存在于猪中性粒细胞、单核细胞、单核细胞表面的独特的和以前未描述的Fc R细胞溶解触发分子复合体和PAM。(C)1995年学术出版社。
G7 and PNK-E mAbs recognize distinct porcine NK cell and granulocyte function-associated molecules that enhance and induce a significant cytolytic response against tumor cell targets through a mechanism of redirected cytotoxicity, The present study shows that the G7 and PNK-E molecules are present on the surface of porcine neutrophils, monocytes, and pulmonary alveolar macrophages as a physically and functionally associated cytolytic trigger molecular complex. Two-color flow cytometric analysis demonstrates that most, if not all, neutrophils are G7 and PNK-E antigen positive, In contrast, monocytes and PAM contain both G7 and PNK-E positive as well as G7 positive and PNK-E negative subpopulations. mAb binding competition experiments indicate that pretreatment of phagocytes with G7 mAb can block subsequent binding by PNK-E mAb, suggesting that these antigens are physically associated on the surface of porcine phagocytes. Fluorescent cocapping experiments utilizing G7 and PNK-E mAbs clearly demonstrate a physical association between G7 and PNK-E antigens present on the surface of porcine neutrophils, Pretreatment of phagocytes with F(ab')(2) fragments of G7 and PNK-E mAbs shows that F(ab')(2) G7 mAb blocks subsequent induction of phagocyte-mediated tumor cell cytotoxicity by whole PNK-E mAb but pretreatment with F(ab')(2) PNK-E does not block subsequent induction of phagocyte-mediated tumor cell cytotoxicity by whole G7 mAb, In addition, pretreatment of neutrophils and mononuclear phagocytes with F(ab')(2) fragments of G7 mAb blocks subsequent whole PNK-E mAb-dependent activation of a phagocytic cell intracellular oxidative burst response but pretreatment with F(ab')(2) PNK-E does not block subsequent G7 mAb-dependent activation of a phagocytic cell intracellular oxidative burst response, These data reinforce a physical and functional association between the G7 and PNK-E molecules, Recent identification of the G7 antigen as the porcine homolog of Fc gamma RIII indicates that the G7 and PNK-E mAbs recognize a unique and previously uncharacterized Fc gamma R cytolytic trigger molecular complex present on the surface of porcine neutrophils,monocytes, and PAM. (C) 1995 Academic Press, Inc.