Intradermal delivery of DNA encoding HCV NS3 and perforin elicits robust cell-mediated immunity in mice and pigs.
Currently, no vaccine is available against hepatitis C virus (HCV), and although DNA vaccines have considerable potential, this has not been realised. Previously, the efficacy of DNA vaccines for human immunodeficiency virus (HIV) and HCV was shown to be enhanced by including the gene for a cytolytic protein, viz. perforin. In this study, we examined the mechanism of cell death by this bicistronic DNA vaccine, which encoded the HCV non-structural protein 3 (NS3) under the control of the CMV promoter and perforin is controlled by the SV40 promoter. Compared with a canonical DNA vaccine and a bicistronic DNA vaccine encoding NS3 and the proapoptotic gene NSP4, the perforin-containing vaccine elicited enhanced cell-mediated immune responses against the NS3 protein in vaccinated mice and pigs, as determined by ELISpot and intracellular cytokine staining, whereas a mouse challenge model suggested that the immunity was CD8(+) T-cell-dependent. The results of the study showed that the inclusion of perforin in the DNA vaccine altered the fate of NS3-positive cells from apoptosis to necrosis, and this resulted in more robust immune responses in mice and pigs, the latter of which represents an accepted large animal model in which to test vaccine efficacy.
Virology Laboratory, Discipline of Surgery, University of Adelaide and Basil Hetzel Institute for Translational Medicine, Adelaide, SA, Australia.
B
B
Grubor-Bauk
Virology Laboratory, Discipline of Surgery, University of Adelaide and Basil Hetzel Institute for Translational Medicine, Adelaide, SA, Australia.
W
W
Yu
Virology Laboratory, Discipline of Surgery, University of Adelaide and Basil Hetzel Institute for Translational Medicine, Adelaide, SA, Australia.
D
D
Wijesundara
Virology Laboratory, Discipline of Surgery, University of Adelaide and Basil Hetzel Institute for Translational Medicine, Adelaide, SA, Australia.
J
J
Gummow
Virology Laboratory, Discipline of Surgery, University of Adelaide and Basil Hetzel Institute for Translational Medicine, Adelaide, SA, Australia.
T
T
Garrod
Killer Cell Biology Laboratory, Cancer Immunology Research, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
A J
AJ
Brennan
Killer Cell Biology Laboratory, Cancer Immunology Research, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
I
I
Voskoboinik
Virology Laboratory, Discipline of Surgery, University of Adelaide and Basil Hetzel Institute for Translational Medicine, Adelaide, SA, Australia.
E J
EJ
Gowans
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D004195:Disease Models, Animal; D058501:Enzyme-Linked Immunospot Assay; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology; D057809:HEK293 Cells; D016174:Hepacivirus; D006801:Humans; D007111:Immunity, Cellular; D007114:Immunization; D008297:Male; D051379:Mice; D054353:Perforin / Q000235:genetics* / Q000276:immunology; D011401:Promoter Regions, Genetic; D013552:Swine; D014118:Toxins, Biological / Q000235:genetics / Q000276:immunology; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000276:immunology; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D004279:DNA, Viral; D006023:Glycoproteins; D054353:Perforin; D014118:Toxins, Biological; D019444:Vaccines, DNA; D017361:Viral Nonstructural Proteins; C081369:NS28 protein, rotavirus; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases