Quantitative evaluation of PpSP15-LmSTI1 fusion gene expression following transfection with an alphavirus-derived self-amplifying mRNA and conventional DNA vaccine platforms

Nastaran Sadat Savar; Thomas Vallet; Masoumeh Azizi; Arash Arashkia; Kenneth Lundstrom; Marco Vignuzzi; Hamid Mahmoudzadeh Niknam

doi:10.1016/j.mcp.2021.101749

Quantitative evaluation of PpSP15-LmSTI1 fusion gene expression following transfection with an alphavirus-derived self-amplifying mRNA and conventional DNA vaccine platforms

Mol Cell Probes. 2021 Oct:59:101749. doi: 10.1016/j.mcp.2021.101749. Epub 2021 Jun 30.

Authors

Nastaran Sadat Savar¹, Thomas Vallet², Masoumeh Azizi³, Arash Arashkia⁴, Kenneth Lundstrom⁵, Marco Vignuzzi⁶, Hamid Mahmoudzadeh Niknam⁷

Affiliations

¹ Immunology Department, Pasteur Institute of Iran, Tehran, Iran.
² Institut Pasteur, Viral Populations and Pathogenesis Unit, Centre National de La Recherche Scientifique UMR, 3569, Paris, France.
³ Molecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
⁴ Virology Department, Pasteur Institute of Iran, Tehran, Iran.
⁵ PanTherapeutics, Switzerland.
⁶ Institut Pasteur, Viral Populations and Pathogenesis Unit, Centre National de La Recherche Scientifique UMR, 3569, Paris, France. Electronic address: marco.vignuzzi@pasteur.fr.
⁷ Immunology Department, Pasteur Institute of Iran, Tehran, Iran. Electronic address: hamid.m.niknam@gmail.com.

PMID: 34214632
DOI: 10.1016/j.mcp.2021.101749

Abstract

New vaccine platforms are crucial to address complex parasitic infections such as cutaneous leishmaniasis. Self-amplifying mRNA (SAM) based vaccines represent the next generation nucleic acid-based platform. In the present study, we compared the expression levels of PpSP15-LmSTI1 fusion gene in BHK-21 cells following transfection with Semliki Forest virus (SFV)-derived SAM, SFV-derived plasmid DNA (pSFV-PD) and conventional plasmid DNA (pcDNA3.1+). PpSP15-LmSTI1 fusion gene expression levels were evaluated at different time points, using quantitative Real-time PCR. All data were validated and normalized by two internal control genes. According to the results, mean values of relative expression were significantly higher for SFV-PD SAM/fusion than pcDNA/fusion and pSFV-PD/fusion at all concentrations and time points. Our results showed that higher levels of PpSp15-LmSTI1 antigen expression could be achieved using a SAM vector than pcDNA and pSFV-PD, making it a valuable and efficient alternative to conventional plasmid DNA-based vaccines against leishmaniasis.

Keywords: Fusion; Gene expression; Plasmid DNA; PpSP15-LmSTI1; Self-amplifying mRNA; Vaccine.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alphavirus* / genetics
Gene Expression
RNA, Messenger / genetics
Transfection
Vaccines, DNA* / genetics

Substances

RNA, Messenger
Vaccines, DNA