Characterization of the cytolytic trigger molecules G7/PNK-E as a molecular complex on the surface of porcine phagocytes

Cell Immunol. 1995 Apr 1;161(2):270-8. doi: 10.1006/cimm.1995.1036.

Abstract

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 re-directed 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 co-capping 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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antigens, Surface / analysis
  • Antigens, Surface / immunology*
  • Binding, Competitive
  • Cytotoxicity, Immunologic
  • Female
  • Flow Cytometry
  • Macrophages, Alveolar / immunology*
  • Male
  • Monocytes / immunology*
  • Neutrophils / immunology*
  • Phagocytosis
  • Receptors, Fc / immunology
  • Receptors, IgG / analysis
  • Receptors, IgG / immunology*
  • Receptors, Immunologic / analysis
  • Receptors, Immunologic / immunology
  • Swine

Substances

  • Antibodies, Monoclonal
  • Antigens, Surface
  • G7 protein, Sus scrofa
  • Receptors, Fc
  • Receptors, IgG
  • Receptors, Immunologic