Study of differentiation from CML into dendritic cell by calcium ionorphore / 中国免疫学杂志

Objective:To explore the methods and condition in which calcium ionorphore(CI) induces the CML cells to differentiate into dendritic cells(DCs).Methods:Mononuclear cells were separated from peripheral blood or bone marrow of CML patients whose WBC counts were more than 30?30~9 L~ -1 when samples were collected,then lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI1640 containing 10%FCS(Fetal calf serum),with or without CI(375 ng/ml) and GM-CSF(200 ng/ml) at 37℃,5%CO_2 humidified atmosphere for 96 h. To evaluate the effect of CI on inducing CML cells to differentiate into DCs,the phenotype of these cells were analyzed by flow cytometry and the morphology change was observed under inverted microscope and electron microscope. Better condition was also explored for DCs differentiation from CML cells under the effect of CI.Results:After 96 h of culture with CI and GM-CSF,the CML cells acquired morphology of mature DCs and significantly up-regulation of CD80,CD86,CD40,CD86 and HLA-DR.Conclusion:CML cells might acquire typical morphology and immunological phenotype of mature DCs when being cultured with CI and GM-CSF.

Upregulation of B7 molecules expression by calcium ionorphore in K562 cells in vitro / 中国免疫学杂志

Objective:To investigate the effect of calcium ionorphore on B7 costimulatory molecules of chronic myeloid leukemia cells line K562.Methods:Well growing K562 cells were cultured in the medium containing calcium ionorphore(375 ng/ml),with K562 cells without CI treatment as control.The cells′ viability and number were calculated by Trypan Blue exclusion at 0,48 and 96 h.Before and after 96 h of cultured,B7-1 and B7-2 expression was assayed by flow cytometry.The proliferation of allogeneic human T cells was measured by MTT colorimetry.Results:B7 costimulatory molecules were abcent or lowly expressed on K562 cells.After 96 h of CI treatment,B7 costimulatory molecules of K562 cells were markedly upregulated and marked activation of allogeneic T cells occurred.No notable morphological change was found during the culture.K562 cells cultured in medium with CI grow slowly than that without CI.Conclusion:B7 costimulatory molecules expression on chronic myeloid leukemia cells line K562 surface was defective.These costimulatory molecules on K562 cells can be upregulated by calcium ionorphore.Calcium ionorphore may inhibit the growth of K562 cells.