Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice
Kunjin virus (KUNV) is an attenuated strain of the severe neurotropic West Nile virus (WNV). The virus has a single-strand positive-sense RNA genome that encodes a polyprotein. Following gene expression, the polyprotein is cleaved into structural proteins for viral packaging and nonstructural protei...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-11-01
|
| Series: | Microorganisms |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2607/8/12/1890 |
| _version_ | 1797546217158213632 |
|---|---|
| author | Pham-Tue-Hung Tran Naveed Asghar Urban Höglund Olivia Larsson Lars Haag Ali Mirazimi Magnus Johansson Wessam Melik |
| author_facet | Pham-Tue-Hung Tran Naveed Asghar Urban Höglund Olivia Larsson Lars Haag Ali Mirazimi Magnus Johansson Wessam Melik |
| author_sort | Pham-Tue-Hung Tran |
| collection | DOAJ |
| description | Kunjin virus (KUNV) is an attenuated strain of the severe neurotropic West Nile virus (WNV). The virus has a single-strand positive-sense RNA genome that encodes a polyprotein. Following gene expression, the polyprotein is cleaved into structural proteins for viral packaging and nonstructural proteins for viral replication and expression. Removal of the structural genes generate subgenomic replicons that maintain replication capacity. Co-expression of these replicons with the viral structural genes produces reporter virus-like particles (RVPs) which infect cells in a single round. In this study, we aimed to develop a system to generate multivalent RVPs based on KUNV to elicit an immune response against different viruses. We selected the Ebola virus (EBOV) glycoprotein (GP) and the matrix protein (VP40) genes, as candidates to be delivered by KUNV RVPs. Initially, we enhanced the production of KUNV RVPs by generating a stable cell line expressing the KUNV packaging system comprising capsid, precursor membrane, and envelope. Transfection of the DNA-based KUNV replicon into this cell line resulted in an enhanced RVP production. The replicon was expressed in the stable cell line to produce the RVPs that allowed the delivery of EBOV GP and VP40 genes into other cells. Finally, we immunized BALB/cN mice with RVPs, resulting in seroconversion for EBOV GP, EBOV VP40, WNV nonstructural protein 1, and WNV E protein. Thus, our study shows that KUNV RVPs may function as a WNV vaccine candidate and RVPs can be used as a gene delivery system in the development of future EBOV vaccines. |
| first_indexed | 2024-03-10T14:26:57Z |
| format | Article |
| id | doaj.art-377d46ced52b46f6931a998e21c821de |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T14:26:57Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-377d46ced52b46f6931a998e21c821de2023-11-20T22:52:30ZengMDPI AGMicroorganisms2076-26072020-11-01812189010.3390/microorganisms8121890Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in MicePham-Tue-Hung Tran0Naveed Asghar1Urban Höglund2Olivia Larsson3Lars Haag4Ali Mirazimi5Magnus Johansson6Wessam Melik7School of Medical Science, Inflammatory Response and Infection Susceptibility Centre (iRiSC), Örebro University, 703 62 Örebro, SwedenSchool of Medical Science, Inflammatory Response and Infection Susceptibility Centre (iRiSC), Örebro University, 703 62 Örebro, SwedenAdlego Biomedical AB, P.O. Box 42, 751 03 Uppsala, SwedenAdlego Biomedical AB, P.O. Box 42, 751 03 Uppsala, SwedenEM Unit (EMil), Department of Laboratory Medicine, Karolinska Institute, 171 77 Solna, SwedenDivision of Clinical Microbiology, Department of Laboratory Medicine, Karolinska Institutet, 141 52 Huddinge, SwedenSchool of Medical Science, Inflammatory Response and Infection Susceptibility Centre (iRiSC), Örebro University, 703 62 Örebro, SwedenSchool of Medical Science, Inflammatory Response and Infection Susceptibility Centre (iRiSC), Örebro University, 703 62 Örebro, SwedenKunjin virus (KUNV) is an attenuated strain of the severe neurotropic West Nile virus (WNV). The virus has a single-strand positive-sense RNA genome that encodes a polyprotein. Following gene expression, the polyprotein is cleaved into structural proteins for viral packaging and nonstructural proteins for viral replication and expression. Removal of the structural genes generate subgenomic replicons that maintain replication capacity. Co-expression of these replicons with the viral structural genes produces reporter virus-like particles (RVPs) which infect cells in a single round. In this study, we aimed to develop a system to generate multivalent RVPs based on KUNV to elicit an immune response against different viruses. We selected the Ebola virus (EBOV) glycoprotein (GP) and the matrix protein (VP40) genes, as candidates to be delivered by KUNV RVPs. Initially, we enhanced the production of KUNV RVPs by generating a stable cell line expressing the KUNV packaging system comprising capsid, precursor membrane, and envelope. Transfection of the DNA-based KUNV replicon into this cell line resulted in an enhanced RVP production. The replicon was expressed in the stable cell line to produce the RVPs that allowed the delivery of EBOV GP and VP40 genes into other cells. Finally, we immunized BALB/cN mice with RVPs, resulting in seroconversion for EBOV GP, EBOV VP40, WNV nonstructural protein 1, and WNV E protein. Thus, our study shows that KUNV RVPs may function as a WNV vaccine candidate and RVPs can be used as a gene delivery system in the development of future EBOV vaccines.https://www.mdpi.com/2076-2607/8/12/1890reporter virus-like particles (RVPs)repliconsKunjin virusEbola virusglycoprotein (GP)matrix protein (VP40) |
| spellingShingle | Pham-Tue-Hung Tran Naveed Asghar Urban Höglund Olivia Larsson Lars Haag Ali Mirazimi Magnus Johansson Wessam Melik Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice Microorganisms reporter virus-like particles (RVPs) replicons Kunjin virus Ebola virus glycoprotein (GP) matrix protein (VP40) |
| title | Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice |
| title_full | Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice |
| title_fullStr | Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice |
| title_full_unstemmed | Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice |
| title_short | Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice |
| title_sort | development of a multivalent kunjin virus reporter virus like particle system inducing seroconversion for ebola and west nile virus proteins in mice |
| topic | reporter virus-like particles (RVPs) replicons Kunjin virus Ebola virus glycoprotein (GP) matrix protein (VP40) |
| url | https://www.mdpi.com/2076-2607/8/12/1890 |
| work_keys_str_mv | AT phamtuehungtran developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice AT naveedasghar developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice AT urbanhoglund developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice AT olivialarsson developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice AT larshaag developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice AT alimirazimi developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice AT magnusjohansson developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice AT wessammelik developmentofamultivalentkunjinvirusreporterviruslikeparticlesysteminducingseroconversionforebolaandwestnilevirusproteinsinmice |