Induction of specific immune response against hepatocellular carcinoma by AFP gene transfected dendritic cells from CD34+ hematopoietic stem cells

    JIA Jun,REN Jun,ZHANG Liª²Wang,ZHANG Hongª²Mei,GUO Xiaoª²Tong,SI Xiaoª²Ming,WANG Jiuª²Hua,CHENGª²Jie

    Department of Oncology,Xijing Hospital,Fourth Military Medical University,Xi¡¯an 710033,China

    ¡¾Abstract¡¿ AIM£º To transfect dendritic cells originating from CD34+ hematopoietic stem cells with AFP gene and to observe the immune response against hepatocellular carcinoma.METHODS£º CD34+hematopoietic stem cells were harvested after mobilization by chemotherapy and Gª²CSF.CD34+ hematopoietic stem cells were induced to differentiate into dendritic cells by cytokine ILª²4,GMª²CSF and TNFª²¦Á.At the fifth day of culture,plasmid pEGFPª²AFP was transfected into dendritic cells using lipofectamine 2000.The immune response triggered by dendritic cells was valued by MTT method.RESULTS£º The dendritic cells originated from CD34+ cells had typical dendritic stick and expressed CD11C,CD80 and CD86.After transfected with AFP gene,the dendritic cells expressed green fluorescence protein and cytochemical assay showed that there were red spots in the cytoplasm of dendritic cells.MTT method detected the cytotoxic Tª²lymphocyte (CTL)reaction against HepG2 cells and showed that the transfected dendritic cells could induce AFP specific immune response.The killing rates of CTL response were respectively (92ª±0 ¡À3ª±5)%,(75ª±5 ¡À4ª±1)% and (72ª±9 ¡À3ª±4)% in AFP gene transfected group,pEGFP transfected group and control group.There was a significant difference between the AFP gene transfected group and the other two groups.CONCLUSION£º AFP gene transfected dendritic cells originating from hematopoietic stem cells can induce specific immune response and stimulate antigen specific CTL response to kill AFPª²expressed hepatocellular carcinoma cells. ......