Decreased survival of human breast cancer cells expressing HER2/neu on in vitro incubation with an anti-HER2/neu antibody fused to C5a or C5a desArg.
Treatment of human epidermal growth factor receptor 2 (HER2/neu)-expressing breast cancer patients with a monoclonal antibody (mAb) directed against HER2/neu improves the outcome of chemotherapy. In cases in which remission is observed, antibody-dependent cell-mediated cytotoxicity (ADCC) seems to be one of the main mechanisms of anti-HER2/neu mAb action, implicating Fc gamma receptors (Fc gamma Rs) in this tumoricidal activity. In vitro and in vivo studies have revealed that anti-HER2/neu-mediated ADCC is mainly accomplished by polymorphonuclear granulocytes (PMN). C5a, a cleavage product of the complement component C5, modulates Fc gamma R expression via upregulation of activating and downregulation of inhibitory Fc gamma Rs. C5a also recruits PMNs to sites of inflammation and increases PMN survival. To enhance the recruitment and activation of C5a receptor-bearing cells into the tumor microenvironment, we developed antibody fusion proteins composed of a human IgG3 anti-HER2/neu antibody genetically fused to C5a [anti-HER2/neu IgG3-(C5a)] or to its derivative, C5a(desArg) [anti-HER2/neu IgG3-(C5a(desArg))]. Both fusion proteins were expressed, properly assembled, and secreted by murine myeloma cells, and displayed chemotactic activity on human PMN. Under comparable conditions, anti-HER2/neu IgG3-(C5a(desArg)) increased the survival of PMN more efficiently than anti-HER2/neu IgG3-(C5a) or C5a(desArg). Surprisingly, incubation of the fusion proteins with breast cancer cells that overexpress HER2/neu (SK-BR-3) induced cell death at a dose at which the anti-HER2/neu IgG3 antibody was innocuous. In the presence of human peripheral blood leukocytes as effector cells, both fusion proteins induced tumor cell death more efficiently than anti-HER2/neu IgG3. These data suggest that anti-HER2/neu IgG3-(C5a) and anti-HER2/neu IgG3-(C5a(desArg)) fusion proteins possess novel properties that could be useful in cancer immunotherapy.
Fuenmayor
Jaheli
J
Laboratorio de Patología Celular y Molecular, Centro de Medicina Experimental, Instituto Venezolano de Investigaciones Científicas, Apartado 20632, Caracas 1020-A, Venezuela. jfuenmay@ivic.gob.ve
Perez-Vazquez
Karin
K
Perez-Witzke
Daniel
D
Penichet
Manuel L
ML
Montano
Ramon F
RF
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000920:Antibody-Dependent Cell Cytotoxicity; D001943:Breast Neoplasms / Q000276:immunology / Q000378:metabolism* / Q000473:pathology*; D045744:Cell Line, Tumor; D002470:Cell Survival; D002634:Chemotaxis, Leukocyte; D015936:Complement C5a / Q000378:metabolism*; D015937:Complement C5a, des-Arginine / Q000378:metabolism*; D005260:Female; D006801:Humans; D018375:Neutrophil Activation; D011485:Protein Binding; D044087:Receptor, Anaphylatoxin C5a / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology / Q000378:metabolism*; D017452:Receptors, IgG / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D015936:Complement C5a; D015937:Complement C5a, des-Arginine; D044087:Receptor, Anaphylatoxin C5a; D018719:Erb-b2 Receptor Tyrosine Kinases; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins