Monoclonal antibody-ricin A chain conjugate selectively cytotoxic for cells bearing the common acute lymphoblastic leukemia antigen.
The toxic subunit of ricin has been conjugated by a disulfide bound to a monoclonal murine antibody (J-5) specific for the common acute lymphoblastic leukemia antigen (CALLA) expressed on human lymphoblastic cells. Both the monoclonal antibody and ricin A chain retained their original biological activity after conjugation. The ricin A chain portion of this conjugate effectively inhibited protein synthesis in a cell-free rabbit reticulocyte lysate system. Its antibody-combining site attached to the membrane of CALLA-bearing cells, whereupon both components of the molecule, antibody and A chain, could be detected using fluorescein-labeled antibody probes directed against either mouse F(ab')2 or ricin A chain. This conjugate proved to be a potent cytotoxin for CALLA-positive Nalm-1 cells growing in vitro and produced 50% inhibition of proliferation at levels of 2 x 10(-10) M. In contrast, CALLA-negative cell lines were unaffected until levels of conjugate approached 10(-6) M. When tested for cytotoxic action on Nalm-1 cells, the composite conjugate molecule was at least 2000 times more effective than J-5 antibody alone, ricin A chain alone, or a noncovalent mixture of these components. Cytotoxicity of the conjugate could be blocked completely by addition of antibodies specific for either mouse immunoglobulin or ricin A chain. Direct comparison of the in vitro cytotoxicity of CALLA-directed univalent Fab versus bivalent F(ab')2 carriers of ricin A chain revealed 70-fold greater efficacy for the latter conjugate. This differential could not be accounted for by differences in A chain activity or binding affinity for the cell membrane but appeared related to the ability of the multivalent agent to induce modulation and internalization of CALLA determinants. No overt toxic effects were noted when the F(ab')2-A chain conjugate was administered i.v. to rabbits at levels as high as 20 mg/kg single injection or 5 mg/kg for 5 days. The unique specificity characteristics and high potency of this cytotoxic conjugate indicate that it has significant therapeutic potential for the treatment of CALLA-bearing human leukemias and lymphomas.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D002460:Cell Line; D002463:Cell Membrane Permeability; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007945:Leukemia, Lymphoid / Q000276:immunology* / Q000473:pathology; D008223:Lymphoma / Q000276:immunology / Q000473:pathology; D009363:Neoplasm Proteins / Q000096:biosynthesis; D010446:Peptide Fragments; D011817:Rabbits; D012276:Ricin / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D007140:Immunoglobulin Fab Fragments; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D012276:Ricin