Augmentation of 17-1A-induced antibody-dependent cellular cytotoxicity by the triple cytokine combination of interferon-alpha, interleukin-2, and interleukin-12.
Previously, interferon-alpha (IFN-alpha), interleukin-2 (IL-2), and interleukin-12 (IL-12) were shown to increase the antibody-dependent cellular cytotoxicity (ADCC) induced by the murine monoclonal antibody 17-1A, which recognizes the tumor-associated antigen EpCAM. In this study, the authors wanted to determine whether the combination of these three cytokines would yield greater cytotoxicity than the single cytokines. For cytotoxicity assessment, a new flow cytometric assay was used that allows the analysis of long-term ADCC exerted by macrophages. Peripheral blood mononuclear cells from healthy donors were used as effector cells against the colorectal carcinoma cell line HT29 at a low effector-to-target ratio of 4.5:1. With this test, the effectiveness of the combinations IL-2 and IFN-alpha, IL-2 and IL-12, and IL-12 and IFN-alpha were compared with each other. The combinations IL-2 plus IL-12 and IFN-alpha plus IL-12 were more potent at the concentrations tested. Furthermore, the triple cytokine combination of IFN-alpha, IL-2, and IL-12 revealed significantly greater ADCC than dual cytokine combinations. Next, CD14+, CD4+, and CD4- cells were isolated by paramagnetic beads and magnetic activated cell sorter (MACS) columns. The CD14+ and CD4- cell populations contained the ADCC effectors. The addition of CD4+ cells to CD14+ or CD4- cells resulted in augmented ADCC, indicating that cooperation between immune cells occurs. These results suggest that multiple cytokine combinations with monoclonal antibodies may be more effective for cancer immunotherapy.
Medizinische Klinik und Poliklinik I, Universität Bonn, Germany.
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D
Flieger
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U
Spengler
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I
Beier
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R
Kleinschmidt
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T
Sauerbruch
I G
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Schmidt-Wolf
D000230:Adenocarcinoma / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D058846:Antibodies, Monoclonal, Murine-Derived; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D015704:CD4 Antigens / Q000276:immunology; D003110:Colonic Neoplasms / Q000276:immunology*; D003601:Cytotoxicity Tests, Immunologic; D005434:Flow Cytometry; D019073:HT29 Cells; D006801:Humans; D016898:Interferon-alpha / Q000276:immunology*; D018664:Interleukin-12 / Q000276:immunology*; D007376:Interleukin-2 / Q000276:immunology*; D018950:Lipopolysaccharide Receptors / Q000276:immunology
D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D015704:CD4 Antigens; D016898:Interferon-alpha; D007376:Interleukin-2; D018950:Lipopolysaccharide Receptors; C400149:Edrecolomab; D018664:Interleukin-12