Biological Characteristics and Mechanism of Killing Tumor of Cytokine-Induced Killer Cells
- VernacularTitle:细胞因子诱导的杀伤细胞生物活性及杀瘤机制
- Author:
miao, LIU
;
xiao-yan, WU
;
run-ming, JIN
- Publication Type:Journal Article
- Keywords:
cytokine-induced killer cell;
immunophenotype;
cytotoxicity;
mechanism of killing tumor
- From:Journal of Applied Clinical Pediatrics
2006;0(15):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the cytotoxicity and mechanism of killing tumor of cytokine-induced killer (CIK) cells in vitro.Methods Mononuclear cells were acquired freshly from bone marrow of children with leukemia,and the cells obstained were induced into dendritic cells by adding granulocyte-macrophage colony-stimulating,IL-4,TNF-? and other cytokines.Lymphocytes cells were isolated freshly from peripheral blood of children with leukemia by Ficoll-Hypaque density centrifugation,and the cells obstained were induced by IFN-?,IL-2 and CD3McAb.The DC cells and CIK cells were co-cultured for 10-25 days,then DC-CIK cells were obtained.Phenotypes of DC-CIK were analyzed by flow cytomery.The cytotoxicity of DC-CIK against a variety of leukemic cell lines was investigated by MTT technique.When treated with mouse-anti-human LFA-1 monoclonal antibody,the expression of GATA-3 and T -bet in the levels of mRNA and protein were mea-sured by using RT-PCR and Western Blot technique,respectively.Results In the first 0-6 days,DC-CIK induced slowly,the proliferation of DC-CIK got 100-fold at the 13th day,cells were rapidly proliferating in the first 13-21 days.The maximum proliferation of DC-CIK reached at the 22nd day.The phenotypes of CD3,CD11a,CD54,HLA-DR were expressed highly; CD3/CD56,CD25,CD28,CD69,FasL were expressed moderately on DC-CIK.The expression of CD16 was not increased.DC-CIK possessed the cytotoxicity against tumor cells of B95,Jhhan and M07e.The effect was stronger to B95,there was no significant difference when the efficiency target ratio was 12.5:1.0 or 25:1,the cytotoxicity reached about 50% and 60%,respectively,against tumor cells of B95.However,it was not obvious to Jhhan and M07e.When the efficiency target ratio was 12.5:1.0 or 25:1,the cytotoxicity reached to 27.21%,25.13%,33.05%,29.72%,respectively,against tumor cells of Jhhan and M07e.When treated with mouse-anti-human LFA-1 monoclonal antibody,the expression of GATA-3 in the level of mRNA was up-regulated(t=3.425,4.523 Pa
