BoHV-4-based vector delivering Ebola virus surface glycoprotein

Abstract

BackgroundEbola virus (EBOV) is a Category A pathogen that is a member of Filoviridae family that causes hemorrhagic fever in humans and non-human primates. Unpredictable and devastating outbreaks of disease have recently occurred in Africa and current immunoprophylaxis and therapies are limited. The main limitation of working with pathogens like EBOV is the need for costly containment. To potentiate further and wider opportunity for EBOV prophylactics and therapies development, innovative approaches are necessary.MethodsIn the present study, an antigen delivery platform based on a recombinant bovine herpesvirus 4 (BoHV-4), delivering a synthetic EBOV glycoprotein (GP) gene sequence, BoHV-4-syEBOVgD106ΔTK, was generated.ResultsEBOV GP was abundantly expressed by BoHV-4-syEBOVgD106ΔTK transduced cells without decreasing viral replication. BoHV-4-syEBOVgD106ΔTK immunized goats produced high titers of anti-EBOV GP antibodies and conferred a long lasting (up to 6 months), detectable antibody response. Furthermore, no evidence of BoHV-4-syEBOVgD106ΔTK viremia and secondary localization was detected in any of the immunized animals.ConclusionsThe BoHV-4-based vector approach described here, represents: an alternative antigen delivery system for vaccination and a proof of principle study for anti-EBOV antibodies generation in goats for potential immunotherapy applications.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-1084-5) contains supplementary material, which is available to authorized users.

Development of a BoHV-4 viral vector expressing tgD of BoHV-1 and evaluation of its immunogenicity in mouse model

“…We used Bovine herpesvirus 4 (BoHV- [30]. For transfection assay and elimination of loxp-BAC-EGFP-Neo-loxp, we used, respectively, human embryonic kidney (HEK293FT; Thermo Fisher Scientific: Catalog number: R70007) and MDBK-cre cells [31].…”

“…To create recombinant BoHV-4 viruses containing the BAC vector (BoHV-4-BAC), we used homologous recombination (HR) in mammalian cells [30].…”

“…To perform recombineering in SW102 bacteria [30], truncated glycoprotein D of BoHV-1 flanked by thymidine kinase of BoHV-4 (TK1-CMV-tgD-Amp-TK2) was amplified from the pC-tgD construct using the TM1 and TM2 primer sets listed in Table 1. These primers contained 50 bp homologous arms to the thymidine kinase gene (access number: AF318573.1) of the BoHV-4 Movar 33/63 strain.…”

“…Finally, the infectious BoHV-4-tgD∆TK viruses were constructed by transfection of extracted BoHV-4-tgD∆TK DNAs into the MDBK cells. The loxp-BAC-CMV-EGFP-loxp cassette was eliminated from the final constructs by virus propagation in MDBK-cre cells [30].…”

Bovine Herpesvirus Type 4 (BoHV-4) Vector Delivering Nucleocapsid Protein of Crimean-Congo Hemorrhagic Fever Virus Induces Comparable Protective Immunity against Lethal Challenge in IFNα/β/γR−/− Mice Models

“…This virus has been successfully explored as a novel herpesviral vector in targeting different viral immunodominant antigens, such as the Ebola surface glycoprotein (BoHV-4-syEBOVgD106ΔTK), Peste des Petits Ruminants virus hemagglutinin (H) (BoHV-4-A-PPRV-H-ΔTK) and various glycoproteins (A29L, M1R and B6R) of Monkeypox virus (BoHV-4-A-CMV-A29LgD 106 ΔTK, BoHV-4-A-EF1α-M1RgD 106 ΔTK and BoHV-4-A-EF1α-B6RgD 106 ΔTK) due to some interesting characteristics. These include easy propagation in cell culture systems, a relatively simple genome with a large capacity for inserting foreign genes, availability of an animal model for investigating its pathogenicity (rabbit) and a lack of documented cell transformation in comparison to other gammaherpesviruses [15,16,17,18,19,20].…”