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.
Flieger
D
D
Medizinische Klinik und Poliklinik I, Universität Bonn, Germany.
Spengler
U
U
Beier
I
I
Kleinschmidt
R
R
Sauerbruch
T
T
Schmidt-Wolf
I G
IG
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