TY - JOUR
T1 - A novel DNA platform designed for vaccine use with high transgene expression and immunogenicity
AU - Babuadze, George Giorgi
AU - Echanove, Jose
AU - Lamarre, Claude
AU - deLaVega, Marc Antoine
AU - Fausther-Bovendo, Hugues
AU - Racine, Trina
AU - M.Gomez, Alejandro
AU - Azizi, Hiva
AU - Wade, Mathew
AU - Kozak, Robert
AU - Kobinger, Gary P.
N1 - Funding Information:
This work was supported by a grant from the Canadian Institutes of Health Research (grant # 143521). RK and GPK were authors who received the grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This study was funded by the Canadian Institute of Health Research grant # PJT-148747 and contribution from the Canadian non-profit GUARD.
Publisher Copyright:
© 2021
PY - 2021/12/3
Y1 - 2021/12/3
N2 - The development of new, low-cost vaccines and effective gene therapies requires accurate delivery and high-level expression of candidate genes. We developed a plasmid vector, pIDV-II, that allows for both easy manipulation and high expression of exogenous genes in mammalian cells. This plasmid is based upon the pVax1 plasmid and shares a common structure with typical mammalian transcription units. It is composed of a chicken β-actin promoter (CAG), followed by an intron and flanked by two restriction sites, and also includes a post-transcriptional regulatory element, followed by a transcriptional termination signal. While the modification of pVax1 elements either decreased eGFP expression levels or had no effect at all, replacement of the promoter, the poly-A signal, deletion of the T7 and AmpR promoters, and inversion of the ORI-Neo/Kan cassette, significantly increased in vitro eGFP expression with the modified plasmid called pIDV-II. To further evaluate our vector, expression levels of three viral antigens were compared in cell lines transfected either with pVax1 or pCAGGS backbones as controls. Higher transgene expression was consistently observed with pIDV-II. The humoral and cellular responses generated in mice immunized with pIDV-II vs pVax1 expressing each viral antigen individually were superior by 2-fold or more as measured by ELISA and ELISPOT assays. Overall these results indicate that pIDV-II induces robust transgene expression, with concomitant improved cellular and humoral immune responses against the transgene of interest over pVax1. The new vector, pIDV-II, offers an additional alternative for DNA based vaccination and gene therapy for animal and human use.
AB - The development of new, low-cost vaccines and effective gene therapies requires accurate delivery and high-level expression of candidate genes. We developed a plasmid vector, pIDV-II, that allows for both easy manipulation and high expression of exogenous genes in mammalian cells. This plasmid is based upon the pVax1 plasmid and shares a common structure with typical mammalian transcription units. It is composed of a chicken β-actin promoter (CAG), followed by an intron and flanked by two restriction sites, and also includes a post-transcriptional regulatory element, followed by a transcriptional termination signal. While the modification of pVax1 elements either decreased eGFP expression levels or had no effect at all, replacement of the promoter, the poly-A signal, deletion of the T7 and AmpR promoters, and inversion of the ORI-Neo/Kan cassette, significantly increased in vitro eGFP expression with the modified plasmid called pIDV-II. To further evaluate our vector, expression levels of three viral antigens were compared in cell lines transfected either with pVax1 or pCAGGS backbones as controls. Higher transgene expression was consistently observed with pIDV-II. The humoral and cellular responses generated in mice immunized with pIDV-II vs pVax1 expressing each viral antigen individually were superior by 2-fold or more as measured by ELISA and ELISPOT assays. Overall these results indicate that pIDV-II induces robust transgene expression, with concomitant improved cellular and humoral immune responses against the transgene of interest over pVax1. The new vector, pIDV-II, offers an additional alternative for DNA based vaccination and gene therapy for animal and human use.
KW - DNA
KW - Immunogenicity
KW - Novel plasmid
KW - Vaccine
KW - Viral antigens
UR - http://www.scopus.com/inward/record.url?scp=85119007496&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85119007496&partnerID=8YFLogxK
U2 - 10.1016/j.vaccine.2021.10.008
DO - 10.1016/j.vaccine.2021.10.008
M3 - Article
C2 - 34774358
AN - SCOPUS:85119007496
SN - 0264-410X
VL - 39
SP - 7175
EP - 7181
JO - Vaccine
JF - Vaccine
IS - 49
ER -