Development of a Multivalent Kunjin Virus Reporter Virus-Like Particle System Inducing Seroconversion for Ebola and West Nile Virus Proteins in Mice

• Published in: Microorganisms (Basel) Vol. 8; no. 12; p. 1890
• Main Authors: Tran, Pham-Tue-Hung, Asghar, Naveed, Höglund, Urban, Larsson, Olivia, Haag, Lars, Mirazimi, Ali, Johansson, Magnus, Melik, Wessam
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 29.11.2020; MDPI
• Subjects: Amino acids; Antibodies; Biologi (Medicinska tillämpningar under 3 och lantbruksvetenskapliga under 4); Biomedical Laboratory Science; Biomedicin; Biomedicine; Biomedicinsk laboratorievetenskap; Biotechnology; Cell culture; Cytomegalovirus; Deoxyribonucleic acid; DNA; Drug delivery systems; Ebola virus; Ebolavirus; Gene expression; Gene transfer; Genes; Genomes; glycoprotein (GP); Glycoproteins; Immune response; Immune system; Immunization; Immunologi (medicinsk under 30110 och lantbruksvetenskaplig under 40302); Infections; Kunjin virus; Matrix protein; matrix protein (VP40); Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Microscopy; Mikrobiologi inom det medicinska området; Molecular Biology; Molecular Cellbiology; Molekylär cellbiologi; Molekylärbiologi; Naturvetenskap; Nonstructural proteins; Packaging; packaging system; Proteins; Replication; replicons; Reporter virus-like particles (RVPs); Ribonucleic acid; RNA; RNA polymerase; Seroconversion; stable cell line; Structural proteins; Transfection; Vaccines; Vector-borne diseases; Viral diseases; Viral envelope proteins; Virus-like particles; Viruses; VP40 protein; West Nile virus; United States > US; Germany; Ebola virus; replicons; Kunjin virus; matrix protein (VP40); glycoprotein (GP); stable cell line; seroconversion; vaccines; packaging system; reporter virus-like particles (RVPs)
• ISSN: 2076-2607; 2076-2607
• DOI: 10.3390/microorganisms8121890

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.