Objective To construct recombinant shuttle plasmid pAdtrack-CMV-Ub-Mep carrying HCV HLA-A2 restricted compound multi-epitopes gene, and transfect to HEK293 cells, to obtain recombinant adenoviruses rAd-Ub-Mep.Methods The compound antigen gene Ub-Mep was cloned into shuttle plasmid pAdTrack-CMV for pAdtrack-CMV-Ub-Mep.The recombinant adenoviruses rAd-Ub-Mep was packed in HEK293 cells by co-transfection with pAdtrack-CMV-Ub-Mep and frame plasmid pAdEasy1.The recombinant adenoviruses rAd-Ub-Mep was identified by PCR and expression of GFP, and the virus titer was determined by TCID50.Results The recombinant adenoviruses rAd-Ub-Mep was confirmed by enzyme cutting, PCR and fluorometric analysis.The virus titer was 1.2×1011cfu/L.Conclusion Success to construct recombinant adenoviruses vaccine, is the base for the study of cellular immunity induced by HCV HLA-A2 restricted compound multi-epitopes gene.
[1]韦三华, 尹文, 雷迎峰, 等.HCV多表位基因重组BCG的筛选及其诱导的免疫应答研究[J].中华微生物学和免疫学杂志, 2007, 27 (11) :1041-1045.
|
[2]Wei SH, Yin W, An QX, et al.A novel HCV vaccine approach using recombinant BCG expressing multi-epitope antigen[J].Archieve of viology, 2008, 153 (6) :1021-1029.
|
[3]韦三华, 董轲, 林芳, 等.HCV HLA-A2限制性复合多表位基因的构建、克隆表达及其免疫特性分析[J].中国免疫学杂志, 2001, 33 (1) :73-75.
|
[4]Houshton M, Abrignani S.Prospects for a vaccine against the hepatitis C virus[J].Nature, 2005, 436 (7053) :961-966.
|
[5]Kawai T, Akira S.Innate immune recognition of viral infection[J].Nat Immunol, 2006, 7 (2) :131-137.
|
[6]Folgori A, Capone S, Ruggeri L, et al.A T-cell HCV vaccine eliciting effective immunity against heterologous virus challenge in chimpanzees[J].Nat Med, 2006, 12 (2) :190-197.
|
[7]Form X, Bukh J, Purcell R H.The challenge of developing a vaccine against hepatitis C virus[J].J Hepatol, 2002, 37 (5) :684-695.
|
[8]Gao M, Wang HP, Wang YN, et al.HCV-NS3 Th1 minigene vaccine based on invariant chain CLIP genetic substitution enhances CD4+Th1 cell responses in vivo[J].Vaccine, 2006, 24 (26) :5491-5497.
|
[9]Ward S, Lauer G, Isba R, et al.Cellular immune responses against hepatitis C virus:the evidence base 2002[J].Clin Exp Immunol, 2002, 128 (2) :195-203.
|
[10]Yu H, Huang H, Xiang J, et al.Dendritic cells pulsed with hepatitis C virus NS3 protein induce immune responses and protection from infection with recombinant vaccinia virus expressing NS3[J].J Gen Virol, 2006, 87 (Pt1) :1-10.
|
[11]Arribillaga L, de Cerio AL, Sarobe P, et al.Vaccination with an adenoviral vector encoding hepatitis C virus (HCV) NS3 protein protects against infection with HCV-recombinant vaccinia virus[J].Vaccine, 2002, (21) :202-210.
|
[12]Hao CQ, Li Gy, Zhou YX, et al.Construction, package and identification of replication-deficient recombinant adenovirus expression vector of HCV C[J].World Chin J Digestol, 2003, 11 (2) :144-147.
|
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