title: Ofatumumab is more efficient than rituximab in lysing B chronic lymphocytic leukemia cells in whole blood and in combination with chemotherapy.
abstract: Ofatumumab (OFA) is a human anti-CD20 Ab approved for treatment of fludarabine-refractory B chronic lymphocytic leukemia (B-CLL). The efficacy of different immunotherapeutic strategies is best investigated in conditions that are as physiologic as possible. We have therefore compared the activity OFA and rituximab (RTX), alone or in combination with chemotherapeutic agents in unmanipulated whole blood assays, using flow cytometry. OFA (10-100 μg/ml) lysed B-CLL targets in whole blood more efficiently and with faster kinetics than RTX, with a mean 56% lysis at 24 h compared with 16%. This activity of OFA was fully complement dependent, as shown by >99% inhibition by anti-C5 Ab eculizumab and a lack of NK cell activation in whole blood. OFA-mediated NK cell activation was blocked by complement. OFA-mediated lysis could be increased an additional 15% by blocking CD55 and CD59 complement inhibitors. Interestingly, OFA-mediated lysis correlated significantly with CD20 expression levels (r(2) = 0.79). OFA showed overlapping dose response curves similar to those for RTX in phagocytosis assays using either human macrophages or neutrophils. However, phagocytosis was inhibited in the presence of serum or whole blood. Finally, combined treatment with mafosfamide and fludarabine showed that these therapeutic drugs are synergistic in B-CLL whole blood assays and show superior activity when combined with OFA compared with RTX. These results confirm in B-CLL samples and in physiologic conditions the superior complement mediated cytotoxicity induced by OFA alone compared with RTX, the lack of NK cell activation, and phagocytosis in these conditions and suggest effective chemoimmunotherapy strategies using this new generation anti-CD20 Ab.
authors.affiliation: Laboratory of Cellular Therapy G. Lanzani, Division of Hematology, Ospedali Riuniti di Bergamo, 24128 Bergamo, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Bologna
authors.forename: Elisa
authors.initials: E
authors.lastname: Gotti
authors.forename: Fabio
authors.initials: F
authors.lastname: Da Roit
authors.forename: Tamara
authors.initials: T
authors.lastname: Intermesoli
authors.forename: Alessandro
authors.initials: A
authors.lastname: Rambaldi
authors.forename: Martino
authors.initials: M
authors.lastname: Introna
authors.forename: Josée
authors.initials: J
authors.lastname: Golay
mesh_terms: D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000097:blood / Q000627:therapeutic use*; D061067:Antibodies, Monoclonal, Humanized; D058846:Antibodies, Monoclonal, Murine-Derived / Q000008:administration & dosage / Q000097:blood / Q000627:therapeutic use*; D018951:Antigens, CD20 / Q000276:immunology; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000008:administration & dosage / Q000097:blood / Q000627:therapeutic use*; D016923:Cell Death / Q000276:immunology; D003167:Complement Activation / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy / Q000276:immunology* / Q000473:pathology*; D000069283:Rituximab; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D058846:Antibodies, Monoclonal, Murine-Derived; D018951:Antigens, CD20; D000069283:Rituximab; C527517:ofatumumab