Cytotoxicity of IFN-gamma-activated dendritic cells to freshly isolated acute myeloid leukemia cells.
- Author:
Jun SHI
1
;
Yi ZHANG
;
Ji-Ying SU
;
Xiao LI
;
Quan PU
;
Kazuma IKEDA
Author Information
1. Department of Hematology, The Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China. shijun7@hotmail.com
- Publication Type:Journal Article
- MeSH:
Acute Disease;
Antigens, CD;
analysis;
B7-2 Antigen;
analysis;
Coculture Techniques;
Cytotoxicity, Immunologic;
drug effects;
immunology;
Dendritic Cells;
drug effects;
immunology;
metabolism;
Fas Ligand Protein;
analysis;
Flow Cytometry;
Humans;
Immunoglobulins;
analysis;
Interferon-gamma;
pharmacology;
Leukemia, Myeloid;
immunology;
pathology;
Membrane Glycoproteins;
analysis;
TNF-Related Apoptosis-Inducing Ligand;
analysis;
Tumor Cells, Cultured;
Tumor Necrosis Factor-alpha;
analysis
- From:
Journal of Experimental Hematology
2005;13(6):1071-1075
- CountryChina
- Language:Chinese
-
Abstract:
To investigate the tumoricidal activity of dendritic cell (DC) stimulated by interferon-gamma (IFN-gamma) against freshly isolated myeloid leukemia cells and its mechanism, the peripheral blood monocytes collected from healthy donors were cocultured with interleukin-4 and granulocyte-macrophage colony-stimulating factor in medium to induce DC for 7 days. After 12 hour culture in the absence or presence of IFN-gamma, the changes of costimulatory molecules were analyzed with flow cytometry. To assay the cytotoxicity of DC against freshly isolated acute myeloid cells, they were cocultured at various effector-to-target ratio for 18 hours, then the percentage of tumoricidal activity was measured with (51)Cr release assay. To explore the mechanism of DC-mediated cytotoxicity, the change of DC surface or intracellular protein expression of Fas ligand (Fas L), TNF-alpha and TNF related apoptosis-inducing ligand (TRAIL) were analyzed with flow cytometry. The results showed that IFN-gamma enhanced cytotoxicity of DC against AML cells was (33.8 +/- 1.6)% at E:T as 20:1, compared with unstimulated DC (P < 0.05); IFN-gamma up-regulated expression of costimulatory molecules of DC surface such as CD86 and CD83; after stimulation with IFN-gamma, expression of intracellular TRAIL of DC was significantly enhanced, but expression of TRAIL on cell surface of DC was low; while the significant changes of Fas L and TNF-alpha expression neither on cell surface or in cells were not observed before or after stimulation with IFN-gamma. It is concluded that DC stimulated by IFN-gamma exhibit tumoricidal activity against AML cells. The cytotoxicity is partially related to maturation of DC and TRAIL inducing apoptosis, but not associated with death domain-independent mechanism of Fas L and TNF-alpha.
