Development of a dendritic cell vaccine encoding multiple cytotoxic T lymphocyte epitopes targeting hepatitis C virus.
The aim of the present study was to develop a dendritic cell (DC) vaccine encoding hepatitis C virus (HCV) multiple cytotoxic T lymphocyte (CTL) epitopes that can stimulate T cell responses in vitro, and can be used for immunization in vivo. DCs were infected with recombinant replication-defective adenoviruses (Ads) expressing 2 HCV sequences fused with green fluorescent protein (GFP) and FLAG tags. One sequence (sequence 1) contained the HCV CTL epitopes, NS4B 1793-1801 and P7 774-782, as well as the HCV Th epitope, NS3 1248-1261. A second sequence (sequence 2) was the positive epitope control which contained HCV core 35-44, core 132-140 and NS3 1248-1261. The efficiency of infection was detected by flow cytometry and the expression of HCV epitopes in the DCs was confirmed by RT-PCR and western blot analysis. Ad infection significantly enhanced DC maturation and interleukin (IL)-12p70 production, resulting in T cell proliferation and increased interferon-γ secretion. The CTLs stimulated by Ad-infected DCs specifically killed Huh7.5 human hepatoma cells. The recombinant Ad-expressing multiple CTL HCV epitopes effectively infected the DCs in vitro and promoted T cell antiviral immune responses, thereby laying the foundation for the development of anti-HCV DC vaccines.
Center of Diagnosis and Treatment for Infectious Diseases of Chinese PLA, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi 710038, PR China.
Yun
Y
Zhou
Futao
F
Zhao
Lin
L
Chen
Li
L
Ma
Yu
Y
Wang
Yu
Y
He
Zhiyuan
Z
Ma
Haili
H
Liu
Yonghong
Y
Guo
Ying
Y
Zhang
Zhiqiang
Z
Yao
Chunqiu
C
Hao
Zhansheng
Z
Jia
D000256:Adenoviridae / Q000276:immunology; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016174:Hepacivirus / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D010641:Phenotype; D011994:Recombinant Proteins / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D014761:Viral Hepatitis Vaccines / Q000276:immunology*
D018984:Epitopes, T-Lymphocyte; D011994:Recombinant Proteins; D014761:Viral Hepatitis Vaccines; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma