title: In vitro cytotoxicity of carcinoma cells with 111In-labeled antibodies to HER-2.
abstract: Antibodies conjugated to radionuclides emitting low-energy electrons, which include Auger electrons and some conversion electrons, were recently shown to efficiently kill cells bearing a high density of the antigen recognized. The primary purpose of this study was to determine if such killing could be obtained with anti-HER-2 antibodies conjugated to (111)In, using the chelator benzyl-diethylenetriaminepentaacetic acid, or (125)I. Target cells were the breast carcinoma SK-BR-3 and the ovarian carcinoma SK-OV-3.ip1. In preliminary experiments, antibody accumulation and catabolism during a 2- to 3-day incubation with antibody was investigated. The level of antibody uptake, in terms of molecules per cell, was high enough such that killing seemed feasible. With an (125)I label, but not an (111)In label, increasing the antibody concentration past a certain point caused a decrease in total antibody accumulation, which might be attributed to effects of antibody binding. To test for cytotoxicity, cells were incubated for 2 days with the labeled antibody, then assayed for colony-forming units with a limiting dilution assay. SK-BR-3 cells were strongly killed ( approximately 3 logs) by antibody 21.1, and 100% kill was obtained by combining two noncompeting antibodies to HER-2 (21.1 and 4D5). SK-OV-3.ip.1 cells were more resistant to killing, but use of the two-antibody mixture produced a surviving fraction of approximately 0.002. (111)In-labeled antibodies to other high-density antigens, epithelial glycoprotein-1 and epithelial glycoprotein-2, also killed these target cells. In contrast, unlabeled antibodies or a nonreactive-labeled antibody produced much less cytotoxicity. The same experiment with an (131)I label (a beta-particle emitter) resulted in much greater levels of nonspecific cytotoxicity and essentially no specific cytotoxicity. This approach may be effective for therapy of micrometastases.
authors.affiliation: Center for Molecular Medicine and Immunology, Belleville, NJ 07109, USA.
authors.forename: Rosana B
authors.initials: RB
authors.lastname: Michel
authors.forename: Philip M
authors.initials: PM
authors.lastname: Andrews
authors.forename: Mary Ellen
authors.initials: ME
authors.lastname: Castillo
authors.forename: M Jules
authors.initials: MJ
authors.lastname: Mattes
mesh_terms: D000906:Antibodies / Q000737:chemistry / Q000276:immunology* / Q000633:toxicity*; D002277:Carcinoma / Q000276:immunology / Q000378:metabolism* / Q000473:pathology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D007205:Indium Radioisotopes; D016499:Radioimmunotherapy; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000906:Antibodies; D007205:Indium Radioisotopes; D018719:Erb-b2 Receptor Tyrosine Kinases