Immunization against hepatitis C virus with a fusion protein containing the extra domain A from fibronectin and the hepatitis C virus NS3 protein.
OBJECTIVE
Vaccination strategies able to induce strong T-cell responses might contribute to eradicate hepatitis C virus (HCV) infection. We previously demonstrated that fusion of an antigen to the extra domain A from fibronectin (EDA) targets the antigen to TLR4-expressing dendritic cells (DC) and improves its immunogenicity. Here, we studied if fusion of EDA with the non-structural HCV protein NS3 might constitute an effective immunogen against HCV.


METHODS
Recombinant NS3 and the fusion protein EDA-NS3 were produced and purified from E. coli, and tested in vitro for their capacity to activate maturation of DC and to favour antigen presentation. HHD transgenic mice expressing the human HLA-A2 molecule were immunized with recombinant proteins in the absence or presence of poly(I:C) and anti-CD40 agonistic antibodies and responses elicited by vaccination were tested in vitro, and in vivo, by their capacity to downregulate intrahepatic expression of HCV-NS3 RNA.


RESULTS
EDA-NS3, but not NS3 alone, upregulated the expression of maturation markers, as well as Delta-like 1 and Delta-like 4 Notch ligands in DC and induced the production of IL-12. Mice immunized with EDA-NS3 had strong and long lasting NS3-specific CD4+ and CD8+ T-cell responses and, in combination with poly(I:C) and anti-CD40, downregulated intrahepatic expression of HCV-NS3 RNA.


CONCLUSIONS
Recombinant EDA-NS3 may be considered for the development of vaccines against HCV infection.
Mansilla
Cristina
C
Division of Hepatology and Gene Therapy, Centre for Applied Medical Research (CIMA), University of Navarra, 31008 Pamplona, Spain.
Gorraiz
Marta
M
Martinez
Marta
M
Casares
Noelia
N
Arribillaga
Laura
L
Rudilla
Francesc
F
Echeverria
Iciar
I
Riezu-Boj
José Ignacio
JI
Sarobe
Pablo
P
Borrás-Cuesta
Francisco
F
Prieto
Jesús
J
Lasarte
Juan José
JJ
D000818:Animals; D000998:Antiviral Agents / Q000627:therapeutic use; D002460:Cell Line; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000378:metabolism; D004195:Disease Models, Animal; D004359:Drug Therapy, Combination; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D005353:Fibronectins / Q000096:biosynthesis / Q000627:therapeutic use*; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006526:Hepatitis C / Q000276:immunology / Q000378:metabolism / Q000517:prevention & control*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011070:Poly I-C / Q000627:therapeutic use; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000627:therapeutic use*; D014760:Viral Fusion Proteins / Q000096:biosynthesis / Q000627:therapeutic use*; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use*; D014765:Viral Vaccines / Q000096:biosynthesis / Q000627:therapeutic use*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000998:Antiviral Agents; D005353:Fibronectins; D015789:HLA-A2 Antigen; D011070:Poly I-C; D012367:RNA, Viral; D011994:Recombinant Proteins; D014760:Viral Fusion Proteins; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases