Modification of monoclonal antibody carbohydrates by oxidation, conjugation, or deoxymannojirimycin does not interfere with antibody effector functions.
Site-specific attachment of metal chelators or cytotoxic agents to the carbohydrate region of monoclonal antibodies results in clinically useful immunoconjugates [Doerr et al. (1991) Ann Surg 214: 118, Wynant et al. (1991) Prostate 18: 229]. Since the capacity of monoclonal antibodies (mAb) to mediate tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) may accentuate the therapeutic effectiveness of immunoconjugates, we determined whether site-specific modification of mAb carbohydrates interfered with these functions. The chemical modifications examined consisted of periodate oxidation and subsequent conjugation to either a peptide linker/chelator (GYK-DTPA) or a cytotoxic drug (doxorubicin adipic dihydrazide). mAb-associated carbohydrates were also modified metabolically by incubating hybridoma cells in the presence of a glucosidase inhibitor deoxymannojirimycin to produce high-mannose antibody. All four forms (unaltered, oxidized, conjugated and high-mannose) of murine mAb OVB-3 mediated tumor cell lysis via CDC. Similarly, equivalent ADCC was observed with native and conjugated forms of mAb OVB-3 and EGFR.1. ADCC was achieved with different murine effector cells such as naive (NS), poly (I*C)- and lipopolysaccharide-stimulated (SS) spleen cells, or Corynebacterium-parvum-elicited peritoneal cells (PEC). All murine effector cell types mediated tumor cell lysis but differed in potency such that PEC > SS > NS. Excellent ADCC activity was also demonstrable by human peripheral blood mononuclear cells with OVB-3-GYK-DTPA and high-mannose OVB-3 mAb. ADCC activity was detectable in vivo: both native and conjugated OVB-3 inhibited growth of OVCAR-3 xenografts in nude mice primed with C. parvum. In conclusion, modification of mAb carbohydrates did not compromise their in vivo or in vitro biological functions. Therefore, combination therapy using immunomodulators to enhance the effector functions of site-specific immunoconjugates could be seriously contemplated.
Awwad
M
M
Department of Biological Research, CYTOGEN Corporation, Princeton, NJ 08540.
Strome
P G
PG
Gilman
S C
SC
Axelrod
H R
HR
D017485:1-Deoxynojirimycin / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology; D002478:Cells, Cultured; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D004317:Doxorubicin / Q000378:metabolism; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D007167:Immunotherapy; D051379:Mice; D008819:Mice, Nude; D008934:Mitogens / Q000378:metabolism; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000188:drug therapy; D009842:Oligopeptides / Q000378:metabolism; D010084:Oxidation-Reduction; D004369:Pentetic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D010504:Periodic Acid / Q000378:metabolism; D013154:Spleen / Q000276:immunology
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D007074:Immunoglobulin G; D008934:Mitogens; D009842:Oligopeptides; D010504:Periodic Acid; D017485:1-Deoxynojirimycin; C062250:glycyl-tyrosyl-lysine-DTPA; D004369:Pentetic Acid; D004317:Doxorubicin; C009288:metaperiodate