title: Improved biodistribution, tumor targeting, and reduced immunogenicity in mice with a gamma 4 variant of Campath-1H.
abstract: Radiolabeled antibodies have shown promise for the treatment of lymphoma and for solid tumor targeting. Campath-1H is a humanized monoclonal antibody that reacts with the CD52 antigen present on human lymphoid and myeloid cells. Campath-1H is a gamma1 (G1) isotype that induces lymphopenia via an Fc-mediated mechanism(s). Isotype switches were engineered, and the resulting antibodies were expressed in NS0 mouse myeloma cells and biosynthetically radiolabeled with [35S]methionine. The forms included G1, G4, and a G4 variant that contained alanine substitutions at (EU numbering) Leu-235, Gly-237, and Glu-318. All isotypes bound antigen equivalently as assessed by target cell binding in vitro. The G4 variant had a greatly reduced capacity to interact with Fc receptor by virtue of reduced binding to THP-1 human myeloid cells and by a 1000-fold increase in EC50 to intermediate antibody-dependent cellular cytotoxicity. The pharmacokinetics of the isotypes were compared in CD-1 (nu/nu) mice bearing an experimental antigen-expressing tumor. The plasma half-life and tumor uptake were increased for the G4 variant. The G4 variant showed significantly less spleen, liver, and bone uptake but similar uptake in the lung, kidney, and stomach and lower tissue-to-blood ratios. Immunogenicity was assessed after repeated monthly administrations of unlabeled antibody in BALB/c mice. A 50% reduction in the incidence of anti-globulin response was observed for the G4 variant. These properties suggest that antibodies with reduced Fc receptor interaction merit additional study as potential targeting vehicles relative to other isotypes for radioimmunotherapy or situations where diminished normal tissue binding contributes to efficacy.
authors.affiliation: Division of Cell Biology, Wellcome Research Laboratories, Research Triangle Park, NC 27709, USA.
authors.forename: J T
authors.initials: JT
authors.lastname: Hutchins
authors.forename: F C
authors.initials: FC
authors.lastname: Kull
authors.forename: J
authors.initials: J
authors.lastname: Bynum
authors.forename: V C
authors.initials: VC
authors.lastname: Knick
authors.forename: L M
authors.initials: LM
authors.lastname: Thurmond
authors.forename: P
authors.initials: P
authors.lastname: Ray
mesh_terms: D000074323:Alemtuzumab; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000493:pharmacokinetics*; D061067:Antibodies, Monoclonal, Humanized; D000912:Antibodies, Neoplasm / Q000096:biosynthesis; D000920:Antibody-Dependent Cell Cytotoxicity; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D005260:Female; D014644:Genetic Variation; D006207:Half-Life; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D016297:Mutagenesis, Site-Directed; D009374:Neoplasms, Experimental / Q000276:immunology* / Q000378:metabolism; D017354:Point Mutation; D011392:Proline; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000493:pharmacokinetics; D012694:Serine; D013329:Structure-Activity Relationship; D014018:Tissue Distribution; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000912:Antibodies, Neoplasm; D011994:Recombinant Proteins; D000074323:Alemtuzumab; D012694:Serine; D011392:Proline