Cell division cyclin 25 homolog C(Cdc25C)is a tumor-associated antigen candidate gene,and this may be used as an effective target in cancer treatment.The present study aims to evaluate the lysis effect of cytotoxic T lymphocytes(CTLs)induced by dendritic cell line DC2.4 overexpressing Cdc25C,and the feasibility of Cdc25C as a component in hepatoma immunotherapy.Methods:The mouse Cdc25C gene was ligated into a lentiviral vector,and transfected into DC2.4 cells.The DC2.4 cell phenotype and cytokine secretion were determined by flow cytometry and ELISA,respectively.CD8+T cells were sorted from the spleens of C57BL/6 mice using a magnetic bead sorting kit obtained from Miltenyi Biotech,Germany,and co-cultured with DC2.4 cells for one week as effector cells.Then,IL-2,granzyme B and perforin were detected in the CTL culture medium by ELISA.Next,time-resolved fluorescence immunoassay was used to detect the immune killing effect of Cdc25C-specific CTLs on target cells.Meanwhile,the effect of blocking MHC-Ⅰ sites on target cells with a monoclonal anti-MHC-Ⅰ antibody was evaluated.Results:The results revealed that Cdc25C could be stably overexpressed in DC2.4 cells by LV-Cdc25C infection.DC2.4 cells transfected with LV-Cdc25C secreted more IL-6,IL-12,TNF-α and IFN-γ,and had higher expression levels of CD40,CD86,CCR7 and MHC-Ⅱ than unaltered DC2.4 cells.The elevated Cdc25C in dendritic cells also further increased the secretion of IL-2,granzyme B and perforin to elicit Cdc25C-specific CTLs,and induced the higher cytotoxicity in Hepal-6 cell lines(P＜0.05),but this had no effect on the target cells when MHC-Ⅰ monoclonal antibodies were blocked.Conclusion:DC2.4 cells transfected with LV-Cdc25C can induce specific CTLs,and result in a strong cellular immune response.The dendritic cells that overexpress Cdc25C may be useful for hepatoma immunotherapy.

Chun-mei LI;Yan-fei LI;Lin TIAN;Qi-hui ZHANG;Fang-yuan ZHENG;Fa-rong MO

School of Basic Medical Science,Guangxi Medical University,Nanning 530021,China;Laboratory of Hepatobiliary and Pancreatic Surgery,Affiliated Hospital of Guilin Medical University,Guilin 541001,China;Guangxi Colleges and Universities Key Laboratory of Human Development and Disease Research,Guangxi Medical University,Nanning 530021,China

当代医学科学(英文)

[1]Chun-mei LI;Yan-fei LI;Lin TIAN;Qi-hui ZHANG;Fang-yuan ZHENG;Fa-rong MO-.Anti-hepatoma Effect of DC2.4 Cells Transfected with Tumor-Associated Antigen Cdc25C In Vitro)[J].当代医学科学(英文),2022(03):491-497

Transfected,Cdc25C,Miltenyi,Biotech,Hepal

hepatoma,Effect,DC2,Cells,Tumor,Associated,Antigen,In,Vitro,division,cyclin,homolog,tumor,associated,antigen,candidate,gene,this,may,used,effective,target,cancer,treatment,present,study,aims,lysis,lymphocytes,CTLs,induced,by,dendritic,overexpressing,feasibility,component,immunotherapy,Methods,mouse,was,ligated,into,lentiviral,vector,transfected,cells,phenotype,cytokine,secretion,were,determined,flow,cytometry,respectively,CD8+T,sorted,from,spleens,C57BL,mice,using,magnetic,bead,sorting,kit,obtained,Germany,cultured,week,effector,Then,granzyme,perforin,detected,medium,Next,resolved,fluorescence,immunoassay,immune,killing,specific,Meanwhile,blocking,MHC,sites,monoclonal,antibody,evaluated,Results,results,revealed,that,could,stably,overexpressed,LV,infection,secreted,more,IFN,had,higher,expression,levels,CD40,CD86,CCR7,than,unaltered,elevated,also,further,increased,elicit,cytotoxicity,lines,but,when,antibodies,blocked,Conclusion,strong,cellular,response,useful

0.458933