Biological Activity of Cytokine-Induced Killer Cells in vitro / 实用儿科临床杂志

ABSTRACT

Objective To investigate the biological activity of cytokine-induced killer(CIK)cells in vitro.Methods Lymphocytes isolated from peripheral blood in leukemic children were induced with interferon-?(IFN-?),anti-CD3 monoclonal antibody(CD3McAb)and interleukin-2(IL-2)and co-cultured with dendritic cells(DC)to generate DC-CIK cells.The morphology and immunophenotype of these cells were determined by electron microscopy and flow cytometry,respectively.Cytotoxicity of DC-CIK cells against leukemia cell lines was measured by the methyl thiazolyl tetrazolium (MTT) assay.Interleukin-12(IL-12),tumor necrosis factor-?(TNF-?)levels released by DC-CIK cells were quantified by enzyme-linked immunosorbent assays.Results Induced DC-CIK cells were regular,round and transparent with variable cell volume and cellular aggregation.At the 0th-4th day,its amplification was very slow,and it increased quickly at the 5th-8th day,it reached its peak amplification at the 9th-10th day,at approximately 100-fold.The main effector cells in this population were CD3+CD8+ cells and CD3+CD56+ cells.DC-CIK cells were cytotoxic to B95 cells,K562 cells and HL-60 cells,with the highest cytotoxicity towards B95 cells.The expression levels of IL-12 and TNF-? in supernatant were very high.Conclusions DC-CIK cells induced with cytokines displayed powerful amplification and strongly killing activities in vitro.It suggested that DC-CIK cells induced with cytokines may play killing activities through Th1 pathway in vitro,as a result of high secretion of Th1 cytokines,such as IL-12 and TNF-?.