Proliferation and identification of dendritic cells from peripheral blood of patients with bladder cancer in vitro / 中华泌尿外科杂志

ABSTRACT

Objective To investigate the proliferation and identification of dendritic cells(DC)de- rived from peripheral blood of patients with bladder cancer in vitro.Methods The mononuclear cells were prepared from peripheral blood of patients with bladder cancer by Ficoll-Hypaque centrifugation method,and were induced by the recombinant cytokines hGM-CSF(50 ng/ml),hlL-4(10 ng/ml)and hTNF-?(50 ng/ ml)for 2 weeks.The growth and morphology of DC were observed through the phase contrast or electron mi- croscope,and their pheuotypes were determined by flow cytometry.The capacity of DC to activate T cell-de- pendent anti-tumor immune responses was tested by MTT method.Results The DC cultured in vitro turned into suspensive growth from adhesive situation on the 6th day,then the number of DC increased con- tinuously and the cells showed the irregular morphologic appearance of DC with veiled edges on the 8th day. Flow cytometry showed that the mature DC expressed high levels of specific markers such as CD_(1a),CD_(83), CD_(86)and HLA-DR.T cells activated by DC showed strong cytotoxicity to bladder cancer cell line BIU87 with a killing rate of(48.8?3.7)%,while the killing rate of T cells which were not activated by DC was(25.7?1.5)%;the difference of the rate between them was significant(P＜0.01). Conclusions The DC can be cultured from peripheral blood of patients with bladder cancer by induction of rhGM-CSF,rhIL-4 and hT- NF-?in vitro.This may lay an experimental foundation for further research on DC vaccine.