Objective To investigate the culture method of cytokines induced killer cells using CD3 monoclonal antibody,IL-2,IFN-γ and IL-1α in vitro.Methods Peripheral blood mononuclear cells (PBMC) were collected by a blood c...Objective To investigate the culture method of cytokines induced killer cells using CD3 monoclonal antibody,IL-2,IFN-γ and IL-1α in vitro.Methods Peripheral blood mononuclear cells (PBMC) were collected by a blood cell separator from 8 patients with refractory lymphoma,then expanded by priming them with recombinant IFN-γ,monoclonal antibody (mAb) to CD3 and human recombinant IL-1α,followed by adding recombinant IL-2 the next day.The CIK cells were counted on d1,4,7,10,13 of incubation,respectively.The phenotypic patterns were characterized before culture and on d13 of culture.Results CIK cells colony formed on d3.On d13,the CIK cells counting reached up to (7-18)×109(mean 12.7×109),multiplying 44-140 times(mean 98 times).Cell survival rate was over 90%.The average percentage of cells expressing CD3+,CD4+,CD8+ and CD3+ CD56+ were also increased from (50.9±3.5)%,(29.9±1.7)%,(41.3±3.2)%,(1.6±0.2)% to (90.2±1.6)%,(40.6±5.5)%,(52.8±4.9)% and (33.1±4.0)%,respectively.Conclusions CIK cells developed by the culture method have high in vitro proliferate rate and tumor-killing capacity.The simplicity of the operation and autogenic source of the cells indicate the method could be applied clinically treating patients with refractory lymphoma.展开更多
文摘Objective To investigate the culture method of cytokines induced killer cells using CD3 monoclonal antibody,IL-2,IFN-γ and IL-1α in vitro.Methods Peripheral blood mononuclear cells (PBMC) were collected by a blood cell separator from 8 patients with refractory lymphoma,then expanded by priming them with recombinant IFN-γ,monoclonal antibody (mAb) to CD3 and human recombinant IL-1α,followed by adding recombinant IL-2 the next day.The CIK cells were counted on d1,4,7,10,13 of incubation,respectively.The phenotypic patterns were characterized before culture and on d13 of culture.Results CIK cells colony formed on d3.On d13,the CIK cells counting reached up to (7-18)×109(mean 12.7×109),multiplying 44-140 times(mean 98 times).Cell survival rate was over 90%.The average percentage of cells expressing CD3+,CD4+,CD8+ and CD3+ CD56+ were also increased from (50.9±3.5)%,(29.9±1.7)%,(41.3±3.2)%,(1.6±0.2)% to (90.2±1.6)%,(40.6±5.5)%,(52.8±4.9)% and (33.1±4.0)%,respectively.Conclusions CIK cells developed by the culture method have high in vitro proliferate rate and tumor-killing capacity.The simplicity of the operation and autogenic source of the cells indicate the method could be applied clinically treating patients with refractory lymphoma.
