[Signal transduction in the differentiation of human peripheral blood mononucleocytes towards dendritic cells induced by calcium ionophore].
OBJECTIVE
To explore the intracellular signal transduction pathway in the differentiation of human peripheral blood mononucleocytes (PBMCs) towards dendritic cells (DCs) induced by calcium ionophore (CI).


METHODS
PBMCs isolated from a healthy donor were cultured with A23187 plus rhGM-CSF, 100 microg/L each. In some experiments, PBMCs were cultured for 30 minutes with W-7 (10 micromol/L), CsA(0.5 mg/L) or KT5926(1 micromol/L) before addition of rhGM-CSF and A23187. After culture for 40 hours, morphological change of the cells were observed under phase contrast microscope; surface markers on treated PBMCs were analyzed by flow cytometry; the proliferation of allogeneic human T cells stimulated by the treated PBMCs was detected by MTT colorimetry.


RESULTS
PBMCs of the healthy donor cocultured with rhGM-CSF plus CI for 40 hours had the typical morphology of DCs, with decreased CD14 expression, and increased CD83, CD80 and CD86 expressions. The proliferation of allogeneic T cells stimulated by PBMCs treated with A23187 plus rhGM-CSF was strengthened. But the morphological changes, surface marker expressions and the ability to enhancing proliferation of allogeneic T cells were inhibited to different degrees by W-7, CsA or KT5926.


CONCLUSIONS
The differentiation of PBMCs towards DCs by CI may be modulated by Ca (2+)/calmodulin and multiple signal transduction pathways downstream.
Wu
Jun
J
Institute of Tumor Molecular Biology, General Hospital of Guangzhou Command, Guangzhou 510010, China. Junwuwan@hotmail.com
Yang
De-mao
DM
Yang
Tai-cheng
TC
Wang
Xiao-huai
XH
Wang
Jie
J
Chen
Zheng-liang
ZL
Fu
Ning
N
D015703:Antigens, CD / Q000378:metabolism; D018122:B7-1 Antigen / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology*; D015220:Calcium Channels / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D007211:Indoles / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000502:physiology*; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D015398:Signal Transduction; D013449:Sulfonamides / Q000494:pharmacology; D000097802:CD83 Antigen
D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D000001:Calcimycin; D015220:Calcium Channels; D002227:Carbazoles; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D007136:Immunoglobulins; D007211:Indoles; D007476:Ionophores; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D013449:Sulfonamides; D000097802:CD83 Antigen; C063769:KT 5926; C017967:W 7