Interleukin 12-based immunotherapy improves the antitumor effectiveness of a low-dose 5-Aza-2'-deoxycitidine treatment in L1210 leukemia and B16F10 melanoma models in mice.
OBJECTIVE
Recent findings indicating that many genes related to cancer development are silenced by an aberrant DNA methylation suggest that inhibitors of this process may be effective cancer therapeutics. In this study we investigated the efficacy of low-dose 5-aza-2'-deoxycitydine (DAC), a methylation inhibitor, with interleukin (IL) 12, one of the most potent cytokines with antitumor activity.


METHODS
Mice inoculated with L1210 leukemia cells or with B16F10 melanoma cells were treated with 7 daily injections of low-dose DAC (0.2 mg/kg) and/or 7 daily doses of IL-12 (100 ng/dose). Scid/scid mice as well as monoclonal antibodies against CD4, CD8, and NK1.1 were used to investigate the mechanisms of the antitumor effects of the combination treatment. The activity of murine lymphocytes was measured with enzyme-linked immunospot and (51)Cr release assays.


RESULTS
Treatment with DAC or IL-12 given alone produced moderate antitumor effects. In both tumor models combined treatment resulted in potentiated antitumor effects and produced 70% long-term survivors among mice inoculated with L1210 cells. The antitumor efficacy of combined treatment was abrogated in scid/scid mice, and after depletion of CD4(+) and CD8(+) T cells. Mice inoculated with B16F10 melanoma cells had significantly delayed tumor growth after combined treatment with DAC and IL-12. Strong antitumor effect correlated with a significant activation of lymph node-derived CD8(+) and CD4(+) cells. Transient neutropenia was observed in mice under treatment of DAC alone, but remarkably this effect was not potentiated by IL-12.


CONCLUSIONS
This study provides the first evidence that antitumor effects of DAC can be strongly potentiated by IL-12 and could be beneficial in an effective low-dose-based antitumor therapy.
Kozar
Katarzyna
K
Department of Immunology, Center of Biostructure, Medical University of Warsaw, 02-004 Warsaw, Poland.
Kamiński
Rafal
R
Switaj
Tomasz
T
Ołdak
Tomasz
T
Machaj
Eugeniusz
E
Wysocki
Piotr J
PJ
Mackiewicz
Andrzej
A
Lasek
Witold
W
Jakóbisiak
Marek
M
Gołab
Jakub
J
D000818:Animals; D000941:Antigens / Q000096:biosynthesis; D000950:Antigens, Ly; D000954:Antigens, Surface; D000964:Antimetabolites, Antineoplastic / Q000627:therapeutic use; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000627:therapeutic use*; D015704:CD4 Antigens / Q000096:biosynthesis; D016827:CD8 Antigens / Q000096:biosynthesis; D045744:Cell Line, Tumor; D002470:Cell Survival; D000077209:Decitabine; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006454:Hemoglobins / Q000096:biosynthesis; D007167:Immunotherapy / Q000379:methods*; D007371:Interferon-gamma / Q000096:biosynthesis / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism / Q000627:therapeutic use*; D015847:Interleukin-4 / Q000378:metabolism; D037181:Lectins, C-Type; D008198:Lymph Nodes / Q000473:pathology; D008214:Lymphocytes / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism* / Q000628:therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D016513:Mice, SCID; D008957:Models, Genetic; D055651:NK Cell Lectin-Like Receptor Subfamily B; D009504:Neutrophils / Q000378:metabolism; D014176:Protein Biosynthesis; D011506:Proteins; D013997:Time Factors
D000941:Antigens; D000950:Antigens, Ly; D000954:Antigens, Surface; D000964:Antimetabolites, Antineoplastic; D000970:Antineoplastic Agents; D015704:CD4 Antigens; D016827:CD8 Antigens; D006454:Hemoglobins; C527211:Klrb1c protein, mouse; D037181:Lectins, C-Type; D055651:NK Cell Lectin-Like Receptor Subfamily B; D011506:Proteins; D018664:Interleukin-12; D015847:Interleukin-4; D000077209:Decitabine; D007371:Interferon-gamma; D001374:Azacitidine