Construction of nervous necrosis virus (NNV) genome-based DNA replicon vectors for the delivery of foreign antigens
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
The advantages of replicon vectors of RNA viruses include a high ability to stimulate innate im-munity and exponential amplification of target mRNA leading to high expression of foreign antigens. The present study aimed to construct a DNA-layered nervous necrosis virus (NNV) replicon vector system in which the capsid protein gene was replaced with a foreign antigen gene and to compare the efficiency of foreign antigen expression between the conventional DNA vaccine vector and the present replicon vector. We presented the first report of a nodavirus DNA replicon-based foreign antigen expression system. Instead of a two-vector system, we devised a one-vector system containing both an NNV RNA-dependent RNA polymerase cassette and a foreign antigen-expressing cassette. This single-vector approach circumvents the issue of low foreign protein expression associated with the low co-transfection efficiency of a two-vector system. Cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 (with the capsid gene ORF replaced with VHSV glyco-protein ORF) exhibited significantly higher transcription of the VHSV glycoprotein gene compared to cells transfected with either a vector without hammerhead ribozyme or a conventional DNA vaccine vector expressing the VHSV glycoprotein. Furthermore, the transcription level of the VHSV glyco-protein in cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 showed a significant increase over time. These results suggest that NNV genome-based DNA replicon vectors have the potential to induce stronger and longer expression of target antigens compared to conventional DNA vaccine vectors.
Abstract
The advantages of replicon vectors of RNA viruses include a high ability to stimulate innate im-munity and exponential amplification of target mRNA leading to high expression of foreign antigens. The present study aimed to construct a DNA-layered nervous necrosis virus (NNV) replicon vector system in which the capsid protein gene was replaced with a foreign antigen gene and to compare the efficiency of foreign antigen expression between the conventional DNA vaccine vector and the present replicon vector. We presented the first report of a nodavirus DNA replicon-based foreign antigen expression system. Instead of a two-vector system, we devised a one-vector system containing both an NNV RNA-dependent RNA polymerase cassette and a foreign antigen-expressing cassette. This single-vector approach circumvents the issue of low foreign protein expression associated with the low co-transfection efficiency of a two-vector system. Cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 (with the capsid gene ORF replaced with VHSV glyco-protein ORF) exhibited significantly higher transcription of the VHSV glycoprotein gene compared to cells transfected with either a vector without hammerhead ribozyme or a conventional DNA vaccine vector expressing the VHSV glycoprotein. Furthermore, the transcription level of the VHSV glyco-protein in cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 showed a significant increase over time. These results suggest that NNV genome-based DNA replicon vectors have the potential to induce stronger and longer expression of target antigens compared to conventional DNA vaccine vectors.
참고문헌 (24)
- Ahne, W., "Spring viremia of carp(SVC)", Dis. Aquat. Organ, 52(3), pp.261~272 (2002)[DOI]
- Atasheva, S., "Enhancement of protein expression by alphavirus replicons by designing self-replicating subgenomic RNAs", P. N. A. S, 111(29), pp.10708~10713 (2014)[DOI]
- Atieh, T., "Haiku: New paradigm for the reverse genetics of emerging RNA viruses", PLoS ONE, 13(2), pp.e0193069~ (2018)[DOI]
- Ball, L. A., "Cellular expression of a functional nodavirus RNA replicon from vaccinia virus vectors", J. Virol, 66(4), pp.2335~2345 (1992)[DOI]
- Berglund, P., "Enhancing immune responses using suicidal DNA vaccines", Nat. Biotechnol, 16, pp.562~565 (1998)[DOI]
- Dominguez, F., "Alphavirus-based replicons demonstrate different interactions with host cells and can be optimized to increase protein expression", J. Virol., 97(11), pp.e0122523~ (2023)[DOI]
- Englezou, P. C., "Self-amplifying replicon RNA delivery to dendritic cells by cationic lipids", Molecular Therapy-Nucleic Acids, 12, pp.118~134 (2018)[DOI]
- Hariharan, M. J., "DNA immunization against herpes simplex virus : enhanced efficacy using a Sindbis virus-based vector", J. Virol, 72, pp.950~958 (1998)[DOI]
- Kim, D. Y., "Enhancement of protein expression by alphavirus replicons by designing self-replicating subgenomic RNAs", Proc. Natl. Acad. Sci. U. S. A, 111(29), pp.10708~10713 (2014)[DOI]
- Kim, S. J., "Juvenile olive flounder immersed in live VHSV at 17 °C and 20°C shows resistance against VHSV infection at 10°C", Virus Res, 273, pp.197738~ (2019)[DOI]
- Lee, K. M., "Replication of heterologous glycoprotein-expressing chimeric recombinant snakehead rhabdoviruses(rSHRVs)and viral hemorrhagic septicemia viruses(rVHSVs)at different temperatures", Virus Res, 297, pp.198392~ (2021)[DOI]
- Leitner, W. W., "Enhancement of tumor-specific immune response with plasmid DNA replicon vectors", Cancer Res, 60, pp.51~55 (2000)
- Ljungberg, K., "Increased immunogenicity of a DNA-launched Venezuelan equine encephalitis virus-based replicon DNA vaccine", J. Virol, 81(24), pp.13412~13423 (2007)[DOI]
- McLoughlin, M. F., "Alphavirus infections in salmonids-a review", J. Fish Dis, 30(9), pp.511~531 (2007)[DOI]
- Näslund, T. I., "Role of innate signalling pathways in the immunogenicity of alphaviral replicon-based vaccines", Virol. J, 8(1), pp.36~ (2011)[DOI]
- Nolden, T., "Reverse genetics in high throughput : rapid generation of complete negative strand RNA virus cDNA clones and recombinant viruses thereof", Sci. Rep, 6, pp.23887~ (2016)[DOI]
- Odegard, A., "Flock house virus : a model system for understanding nonenveloped virus entry and membrane penetration", Curr. Top. Microbiol. Immunol, 343, pp.1~22 (2010)[DOI]
- Panzarin, V., "In vitro study of the replication capacity of the RGNNV and the SJNNV betanodavirus genotypes and their natural reassortants in response to temperature", Vet. Res, 45(1), pp.56~ (2014)[DOI]
- Reap, E. A., "Development and preclinical evaluation of an alphavirus replicon particle vaccine for cytomegalovirus", Vaccine, 25(42), pp.7441~7449 (2007)[DOI]
- Rouxel, R. N., "Efficient co-replication of defective novirhabdovirus", Viruses, 8(3), pp.69~ (2016)[DOI]
- Toffan, A., "Water temperature affects pathogenicity of different betanodavirus genotypes in experimentally challenged Dicentrarchus labrax", Dis. Aquat. Org, 119(3), pp.231~238 (2016)[DOI]
- Tonkin, D. R., "Alphavirus replicon-based enhancement of mucosal and systemic immunity is linked to the innate response generated by primary immunization", Vaccine, 28(18), pp.3238~3246 (2010)[DOI]
- Venter, P. A., "Recent insights into the biology and biomedical applications of Flock House virus", Cell. Mol. Life Sci, 65(17), pp.2675~ (2008)[DOI]
- Zhou, Y., "Bipartite viral RNA genome heterodimerization influences genome packaging and virion thermostability", J. Virol., 98(3), pp.e0182023~ (2024)[DOI]