[Cytotoxicity induced by HBsAg gene modified-dendritic cells against hepatocellular carcinoma cell HepG2.2.15].
OBJECTIVE
Up to now, there is no efficient immunotherapy for hepatocellular carcinoma (HCC). Dendritic cell (DC) vaccine could be a potential tool for HCC immunotherapy. This study was to evaluate the effect of dendritic cells (DCs) transfected with recombinant plasmid bearing hepatitis B virus surface antigen (HBsAg) gene, and the capability of generating specific cytotoxic T lymphocytes (CTL) response against HepG2.2.15 in vitro, which were induced by genetically modified DCs.


METHODS
After cultured for 5 days, the DCs were transfected with pCR3.1-S by liposome. The HBsAg gene expression on pCR3.1-transfected DCs was identified by Western blot analysis, and immunofluorescence methods. The cytotoxicity against HepG2.2.15, which were induced by DCs, was tested by MTT assay.


RESULTS
DCs up-regulated the expression of CD1a (55.0%), CD11c (98.6%), CD86 (86.1%), CD80 (66.1%), and HLA-DR (88.9%) after cultured for 5 days. Indirect immunofluorescence, and Western blot analysis showed that HBsAg gene was expressed on transfected DCs. The death rates of HepG2.2.15 cells induced by DCs transfected with pCR3.1-S were (52.3+/-2.8)% (E:T=5:1), (64.6+/-2.4)% (10:1), (78.8+/-2.6) (20:1), (82.1+/-2.4)% (40:1), while the pCR3.1- transfected and non-transfected DCs only induced relatively lower cytotoxicity (P< 0.05, n=4).


CONCLUSIONS
DCs transfected with recombined plasmid expressed HBsAg efficiently, and the genetically modified DCs evoke a higher CTL response in vitro.
Department of Oncology, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, 710032, PR China.
Jing-Yue
JY
Yang
Jun
J
Ren
Jun
J
Bai
Du-Hu
DH
Liu
Li
L
Fan
Xiao-Ming
XM
Si
Zeng-Hui
ZH
Teng
Wen-Tao
WT
Yang
D039521:CD11c Antigen / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000276:immunology / Q000473:pathology* / Q000821:virology; D016923:Cell Death; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D005814:Genes, Viral; D006684:HLA-DR Antigens / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008113:Liver Neoplasms / Q000276:immunology / Q000473:pathology* / Q000821:virology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection
D039521:CD11c Antigen; D006684:HLA-DR Antigens; D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins