Optimization and Limitation of Calcium Ionophore to Generate DCs from Acute Myeloid Leukemic Cells
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Purpose: Calcium ionophore (CI) is used to generate dendritic cells (DCs) from progenitor cells, monocytes, or leukemic cells. The aim of this study was to determine the optimal dose of CI and the appropriate length of cell culture required for acute myeloid leukemia (AML) cells and to evaluate the limitations associated with CI. Materials and Methods: To generate leukemic DCs, leukemic cells (4×106 cells) from six AML patients were cultured with various concentrations of CI and/or IL-4 for 1, 2 or 3 days. Results: Potent leukemic DCs were successfully generated from all AML patients, with an average number of 1.2×106 cells produced in the presence of CI (270 ng/ml) for 2 days. Several surface molecules were clearly upregulated in AML cells supplemented with CI and IL-4, but not CD11c. Leukemic DCs cultured with CI had a higher allogeneic T cell stimulatory capacity than untreated AML cells, but the addition of IL-4 did not augment the MLR activity of these cells. AML cells cultured with CI in the presence or absence of IL-4 showed increased levels of apoptosis in comparison to primary cultures of AML cells. Conclusion: Although CI appears to be advantageous in terms of time and cost effectiveness, the results of the present study suggest that the marked induction of apoptosis by CI limits its application to the generation of DCs from AML cells. (Cancer Res Treat. 2007;39:175-180)
Abstract
Purpose: Calcium ionophore (CI) is used to generate dendritic cells (DCs) from progenitor cells, monocytes, or leukemic cells. The aim of this study was to determine the optimal dose of CI and the appropriate length of cell culture required for acute myeloid leukemia (AML) cells and to evaluate the limitations associated with CI. Materials and Methods: To generate leukemic DCs, leukemic cells (4×106 cells) from six AML patients were cultured with various concentrations of CI and/or IL-4 for 1, 2 or 3 days. Results: Potent leukemic DCs were successfully generated from all AML patients, with an average number of 1.2×106 cells produced in the presence of CI (270 ng/ml) for 2 days. Several surface molecules were clearly upregulated in AML cells supplemented with CI and IL-4, but not CD11c. Leukemic DCs cultured with CI had a higher allogeneic T cell stimulatory capacity than untreated AML cells, but the addition of IL-4 did not augment the MLR activity of these cells. AML cells cultured with CI in the presence or absence of IL-4 showed increased levels of apoptosis in comparison to primary cultures of AML cells. Conclusion: Although CI appears to be advantageous in terms of time and cost effectiveness, the results of the present study suggest that the marked induction of apoptosis by CI limits its application to the generation of DCs from AML cells. (Cancer Res Treat. 2007;39:175-180)
참고문헌 (25)
- Lowenberg B, "Acute myeloid leukemia", 341, pp.1051~1062 (1999)
- Buggins AG, "Effect of costimulation and the microenvironment on antigen presentation by leukemic cells", 94, pp.3479~3490 (1999)
- Notter M, "immunoglobulin G fusion protein to acute myeloid leukemia blasts increases their costimulatory activity for autologous remission T cells", 97, pp.3138~3145 (2001)
- Coppock DL, "Kern J. Identification of a malignant counterpart of the monocyte- dendritic cell progenitor in an acute myeloid leukemia. Blood. 1994;84", pp.3054~62
- Hart DN, "Dendritic cells: unique leukocyte populations which control the primary immune response", 90, pp.3245~3287 (1997)
- Banchereau J, "Dendritic cells and the control of immunity", 392, pp.245~252 (1998)
- Harrison BD, "Stimulation of autologous proliferative and cytotoxic T-cell responses by leukemic dendritic cells derived from blast cells in acute myeloid leukemia", 97, pp.2764~2771 (2001)
- Lee JJ, "The generation of leukemic dendritic cells from acute myeloid leukemia cells is potentiated by the addition of CD40L at the terminal maturation stage", 19, pp.130~136 (2004)
- Reid DC, "Dendritic cells and immunotherapy for malignant disease", 112, pp.874~887 (2001)
- Waclavicek M, "Calcium ionophore: a single reagent for the differentiation of primary human acute myelogenous leukaemia cells towards dendritic cells", 114, pp.466~473 (2001)
- Galea-Lauri J, "Immunological weapons against acute myeloid leukemia", 107, pp.20~27 (2002)
- Lee JJ, "Induction of leukemic-cell-specific cytotoxic T lymphocytes by autologous monocyte-derived dendritic cells presenting leukemic cell antigens", 21, pp.188~194 (2006)
- Czerniecki BJ, "Calcium ionophore-treated peripheral blood monocytes and dendritic cells rapidly display characteristics of activated dendritic cells", 159, pp.3823~3837 (1997)
- Engels FH, "Calcium signaling induces acquisition of dendritic cell characteristics in chronic myelogenous leukemia myeloid progenitor cells", 96, pp.10332~10337 (1999)
- Koski GK, "Calcium ionophore-treated myeloid cells acquire many dendritic cell characteristics independent of prior differentiation state, transformation status, or sensitivity to biologic agents", 94, pp.1359~1371 (1999)
- Westers TM, "Rapid generation of antigen- presenting cells from leukaemic blasts in acute myeloid leukaemia", 52, pp.17~27 (2003)
- Slukvin II, "Directed differentiation of human embryonic stem cells into functional dendritic cells through the myeloid pathway", 176, pp.2924~2932 (2006)
- Lee JJ, "Generation of cytotoxic donor CD8+ T cells against relapsing leukemic cells following allogeneic transplantation by stimulation with leukemic cell- or leukemic lysate pulsed donor cell-derived dendritic cells", 28, pp.517~524 (2004)
- Choi BH, "Optimization of the concentration of autologous serum for generation of leukemic dendritic cells from acute myeloid leukemia cells for clinical immunotherapy", 21, pp.233~240 (2006)
- Kang HK, "Down-regulation of cellular VEGF levels induces differentiation of leukemic cells to functional leukemic-dendritic cells in acute myeloid leukemia", 47, pp.2224~2233 (2006)
- Davis TA, "Phorbol esters induce differentiation of human CD34+ hemopoietic progenitors to dendritic cells: evidence for protein kinase C-mediated signaling", 160, pp.3689~3697 (1998)
- Koski GK, "Calcium mobilization in human myeloid cells results in acquisition of individual dendritic cell-like characteristics through discrete signaling pathways", 163, pp.82~92 (1999)
- Engels FH, "Calcium ionophore activation of chronic myelogenous leukemia progenitor cells into dendritic cells is mediated by calcineurin phosphatase", 24, pp.795~804 (2000)
- Li Q, "Protein kinase C blockade inhibits differentiation of myeloid blasts into dendritic cells by calcium ionophore A23187", 81, pp.131~137 (2005)
- Kharfan-Dabaja M, "Differentiation of acute and chronic myeloid leukemic blasts into the dendritic cell lineage: analysis of various differentiation-inducing signal", 54, pp.25~36 (2005)