Targeting of complement to tumor cells by heteroconjugates composed of antibodies and of the complement component C3b.
Tumor cells have adapted several strategies which permit them to grow in an immunologically hostile environment. The C system can potentially destroy these cells; however, its action needs to be specifically potentiated on the surface of the tumor cells. To this end, a heteroconjugate composed of a mouse mAb and of the human C3b C component has been generated by using the heterobifunctional reagent N-succinimidyl-3-(2-pyridyldithio)propionate. The two mAb which were used in this study are V1-10 and TIB219 which bind to the human and mouse transferrin receptors, respectively. The mAb-C3b conjugates were purified by gel filtration and were each composed of one mAb and one C3b. They bound to the human K562 and HL60 or mouse ALB1 cell lines and amplified the killing of these cells by C from 10 to 15% to 70 to 100%. Fresh normal human or mouse sera were used as a source of C. The mAb-C3b conjugates activated primarily the alternative pathway of C since only C3 and factor B but not C4 were cleaved in the sera. After disulfide-linking to the mAb, the C3b became highly resistant to inactivation by factors H and I, probably due to its reduced factor H binding capacity. On the other hand, the conjugated C3b bound factor B better than free C3b and produced more C3 convertases which expressed increased stability. These results suggest that mAb-C3b conjugates may serve as an effective tool for the specific activation of the cytolytic C system on selected cells. As such, they may be used in vitro or in vivo to target the autologous C to tumor cells or to lymphocytes and may promote tumor immunotherapy.
Reiter
Y
Y
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Fishelson
Z
Z
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage*; D050577:Complement C3-C5 Convertases / Q000494:pharmacology; D003179:Complement C3b / Q000008:administration & dosage* / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism / Q000494:pharmacology; D017242:Complement Factor H; D003170:Complement Pathway, Alternative; D003602:Cytotoxicity, Immunologic; D004546:Elapid Venoms / Q000008:administration & dosage; D006801:Humans; D007167:Immunotherapy; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009369:Neoplasms / Q000276:immunology* / Q000628:therapy; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D004546:Elapid Venoms; C016310:cobra venom factor; D003179:Complement C3b; D017242:Complement Factor H; D050577:Complement C3-C5 Convertases