title: Investigation of the mechanism of drug-induced autoimmune hemolytic anemia in cynomolgus monkeys elicited by a repeated-dose of a humanized monoclonal antibody drug.
abstract: We investigated the mechanism of hemolytic anemia detected in a repeated-dose toxicity study using cynomolgus monkeys that were treated with a humanized antibody drug. This drug was an IgG1 monoclonal antibody (MoAb) that binds to the human HM1.24 antigen named anti-HM1.24 MoAb. The presence of the HM1.24 antigen on the erythrocyte membranes and the erythrocyte agglutination following the addition of anti-HM1.24 MoAb was examined. In addition, an indirect Coombs' test, a hemolysis assay and the measurement of anti-single stranded-DNA antibodies were performed using test animal serum or plasma. The specific binding of FITC- and 125I-labeled anti-HM1.24 MoAb to the erythrocyte membrane was not observed. HM1.24 antigen was not identified on the erythrocyte membranes. However, a high concentration (more than 713 microg/mL) of anti-HM1.24 MoAb hemagglutinated the erythrocyte suspensions. The cause of this agglutination was unclear, but it is assumed that the non-specific binding and/or adhesion caused the direct agglutination. In the examination using test serum from the anemic monkeys, a positive reaction in the indirect Coombs' test was noted. Moreover, in these Coombs' test-positive animals, the production of anti-single stranded-DNA antibodies was sequentially increased. In the female monkey sacrificed in extremis due to severe anemia, an in vitro hemolytic reaction was detected attributable to complement activation. From these results, the hemolytic anemia detected in the repeated-dose toxicity study was diagnosed as a drug-induced autoimmune hemolytic anemia (AIHA) and the primary cause was assumed to be production of IgG class anti-erythrocyte autoantibodies.
authors.affiliation: Fuji Gotemba Laboratories, Chugai Pharmaceutical Co., Ltd., 1-135 Komakado, Gotemba-shi, Shizuoka, 412-8513, Japan.
authors.forename: Kazuto
authors.initials: K
authors.lastname: Watanabe
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Watanabe
authors.forename: Akifumi
authors.initials: A
authors.lastname: Shioda
authors.forename: Keiji
authors.initials: K
authors.lastname: Mizoguchi
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Sugimoto
authors.forename: Keiji
authors.initials: K
authors.lastname: Terao
mesh_terms: D000744:Anemia, Hemolytic, Autoimmune / Q000139:chemically induced* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000633:toxicity*; D000954:Antigens, Surface / Q000276:immunology; D001323:Autoantibodies / Q000032:analysis; D003298:Coombs Test; D004277:DNA, Single-Stranded / Q000276:immunology; D004305:Dose-Response Relationship, Drug; D004903:Erythrocyte Aggregation / Q000187:drug effects / Q000276:immunology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000276:immunology; D004912:Erythrocytes / Q000187:drug effects / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D006461:Hemolysis / Q000187:drug effects; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism / Q000633:toxicity; D008252:Macaca fascicularis; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000378:metabolism; D018675:Toxicity Tests
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D001323:Autoantibodies; D004277:DNA, Single-Stranded; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins