The Killing Effects Of Immune Cells Induced By Leukemia Stem Cell-derived Dendritic Cells On Leukemia Stem Cells From The Same Source

ObjectivesTo investigate the mature immunophenotypes,changes of DC and CIK before and after co-cultured with each other;and to investigate the killing effects of stem cell-derived Dendritic Cells(DC)co-cultured with cytokine induced killer(CIK)on Leukemia Stem Cells(LSCs)from the same source.MethodsBone marrow cells were isolated from CML-AP donors,mononuclear cells were separated by Ficoll-Hypaque.Marked the cells with CD34+CD38-CD44+,leukemia stem cells were sorted by flow cytometry.DC were amplified and induced by cytokines.Cell morphology was observed everyday.The expressions of DC's markers typical?P-gp resistance protein were detected by flow cytometry.The expressions of bcr/abl fusion gene of leukemia stem cells and DC were detected by fluorescence in situ hybridization(FISH).The mononuclear cells from the same doner were induced to CIK.The expressions of CIK's markers typical were detected by flow cytometry.DC and CIK were co-cultured acording to the ratio of 1:10.The expressions of markers typical were detected by flow cytometry a week later.DC and CIK were co-cultured to determined the killing effects.The cytotoxicity activity was measured by lactate dehydrogenase release assay.Results1.DC with typical CML-LSCs,festures.Leukemia stem cell-derived DC express P-gp,the positive expression rate was(60.61±12.38)%.The expression rate of bcr/abl fusion gene in DC was(48.42±20.69)%.DC express P-gp,and bcr/abl fusion gene was positive.Therefore,DC with typical CML-LSCs' festures.2.The expression of DC's immunophenotypes after co-cultured with CIK.DC's immunophenotypes after co-cultured with CIK,the expression rates of CD40,CD80,CD83 and HLA-DR were increased from(41.28 ± 10.76)%,(46.35±13.10)%,(32.41 ±4.30)%,(50.28±9.93)%to(77.79±11.98)%,(81.50±9.45)%,(82.76±6.20)%,(70.48±11.01)%,the differences were significant(P<0.05).3.The expression of CIK's immunophenotypes after co-cultured with DC.CIK's immunophenotypes after co-cultured with DC,the mature immunophenotypes of CD3,CD3CD8,CD3CD56 were increased from(81.65±8.58)%,(51.07±7.20)%,(33.25±6.07)%to(89.98±6.57)%,(79.74±8.22)%,(60.37±9.54)%,the differences were significant(P<0.05 or P<0.01).4.The killing effects in different effector-target ratio.The co-cultered cells were effector cells,LSCs were targe cells.The killing effects in 10:1,20:1,40:1 effector-target ratio were(11.15±3.94)%,(31.13±12.01)%,(59.13±12.82)%.There were significant differences of the killing effects in the three groups(P<0.01).The killing effect increased with effector-target ratio,and there were significant differences of the killing effects between each two groups(P<0.01).ConclusionLeukemia stem cell-derived DC's functions were normal,and killing effects can be found when they were co-cultured with CIK.