[Research on A23187 inducing HL-60 cells to differentiate into dendritic cells].
OBJECTIVE
To investigate the method for calcium mobilization inducing rapidly to produce the dendritic cells (DC) from myeloid leukemia cells.


METHODS
HL-60 cells were cultured with calcium ionophore A23187 (45-720 ng/mL) or plus rhIFN-gamma (1000 U/mL) for 20-96 hours. The morphologic features of cells were observed under inverted microscope and scanning electron microscope, when the cell phenotypes of HL-60 treated with A23187 were determined by flow cytometry. The proliferation of allogeneic human T cells was tested by mixed lymphocyte reaction (Allo-MLR).


RESULTS
After treated with A23187 (180 ng/mL) or plus rhIFN-gamma (1000 U/mL) for 20 hours, some of HL-60 cells were found to have the dendritic appearance on cell surface; and the CD83 as a characteristic marker of mature DCs, co-stimulating molecules CD80 and CD86 got an up-regulated expression. When HL-60 cells were cultured with A23187 for 48 hours, the expression of CD83 began going down. A large number of cells with typical dendritic appearance were observed after cultured with A23187 for 72 hours, and the expressions of CD80 and CD86 were up-regulated continuously. Allo-MLR revealed that DCs derived from HL-60 cells treated with A23187 or plus rhIFN-gamma could stimulate the proliferation of allogeneic human T cells.


CONCLUSIONS
The calcium ionophore A23187 can induce the HL-60 cells into mature DCs. This study suggests that calcium mobilization for cell differentiation may be an alternative way to rapidly acquire DCs.
Yang
Kun
K
Department of Pediatrics, West China Second Hospital, Sichuang University, Chengdu 610041, China.
Li
Qiang
Q
Zhao
Ya-Ning
YN
D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D000001:Calcimycin / Q000276:immunology / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D013997:Time Factors
D000954:Antigens, Surface; D000001:Calcimycin; D007371:Interferon-gamma; D002118:Calcium