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      Safety studies with the oral rabies virus vaccine strain SPBN GASGAS in the small Indian mongoose ( Herpestes auropunctatus)

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          Abstract

          Background

          Oral vaccination of the small Indian mongoose against rabies has been suggested as a potential tool to eliminate mongoose-mediated rabies on several Caribbean islands. A recently developed oral rabies virus vaccine strain, SPBN GASGAS, has already been shown to be efficacious in this reservoir species. Since, all available oral rabies vaccines are based on replication-competent viruses and vaccine baits are distributed unsupervised in the environment, enhanced safety standards for such vaccine types are required.

          Results

          The results of safety studies, including overdose, repeated doses, dissemination and different routes of administration, in the target species are presented. It was shown that the construct was apathogenic, irrespective of dose and route of administration. Even when it was inoculated directly in the brain, it did not induce rabies infection. Furthermore, the vaccine strain did not spread within the target species after direct oral instillation beyond the site of entry.

          Conclusion

          The vaccine strain SPBN GASGAS meets the safety requirements for live rabies virus vaccines in this target species, the small Indian mongoose.

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          Most cited references36

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          Development of a fluorescent antibody virus neutralisation test (FAVN test) for the quantitation of rabies-neutralising antibody.

          A microtest named the FAVN test (fluorescent antibody virus neutralisation test), which is an adaptation of the original rapid fluorescent focus inhibition test (RFFIT) has been developed and evaluated. One hundred percent specificity was estimated using 414 sera from dogs sampled in rabies-free areas or from non-vaccinated animals. The accuracy as determined by the agreement between observed and expected values using sera of known titres was satisfactory. Serum samples from unvaccinated and vaccinated dogs (using sera with titres near 0.5 IU/ml) were assayed for rabies antibody by the FAVN test, the RFFIT and the mouse neutralisation test (MNT): comparative results obtained on the same sera with the three tests showed good agreement. Furthermore, distinguishing negative sera from positive sera with low titres is much easier with the FAVN test than with the RFFIT.
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            Infectious rabies viruses from cloned cDNA.

            The generation of infectious rabies virus (RV), a non-segmented negative-stranded RNA virus of the Rhabdoviridae family, entirely from cloned cDNA is described. Simultaneous intracellular expression of genetically marked full-length RV antigenome-like T7 RNA polymerase transcripts and RV N, P and L proteins from transfected plasmids resulted in formation of transcriptionally active nucleocapsids and subsequent assembly and budding of infectious rabies virions. In addition to authentic RV, two novel infectious RVs characterized by predicted transcription patterns were recovered from modified cDNA. Deletion of the entire non-translated pseudogene region, which is conserved in all naturally occurring RVs, did not impair propagation of the resulting virus in cell culture. This indicates that non-essential genetic material might be present in the genomes of non-segmented RNA viruses. The introduction of a functional extra cistron border into the genome of another virus resulted in the transcription of an additional polyadenylated mRNA containing pseudogene sequences. The possibility of manipulating the RV genome by recombinant DNA techniques using the described procedure--potentially applicable also for other negative-stranded viruses--greatly facilitates the investigation of RV genetics, virus-host interactions and rabies pathogenesis and provides a tool for the design of new generations of live vaccines.
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              Overexpression of the rabies virus glycoprotein results in enhancement of apoptosis and antiviral immune response.

              A recombinant rabies virus (RV) carrying two identical glycoprotein (G) genes (SPBNGA-GA) was constructed and used to determine the effect of RV G overexpression on cell viability and immunity. Immunoprecipitation analysis and flow cytometry showed that tissue culture cells infected with SPBNGA-GA produced, on average, twice as much RV G as cells infected with RV carrying only a single RV G gene (SPBNGA). The overexpression of RV G in SPBNGA-GA-infected NA cells was paralleled by a significant increase in caspase 3 activity followed by a marked decrease in mitochondrial respiration, neither of which was observed in SPBNGA-infected cells. Furthermore, fluorescence staining and confocal microscopy revealed an increased extent of apoptosis and markedly reduced neurofilament and F actin in SPBNGA-GA-infected primary neuron cultures compared with neuronal cells infected with SPBNGA, supporting the concept that RV G or motifs of the RV G gene trigger the apoptosis cascade. Mice immunized with SPBNGA-GA showed substantially higher antibody titers against the RV G and against the nucleoprotein than SPBNGA-immunized mice, suggesting that the speed or extent of apoptosis directly determines the magnitude of the antibody response.
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                Author and article information

                Contributors
                Steffen.Ortmann@idt-biologika.de
                Ad.Vos@idt-biologika.de
                Antje.Kretzschmar@idt-biologika.de
                Nomusa.Walther@idt-biologika.de
                Christiane.Kaiser@idt-biologika.de
                Conrad.Freuling@fli.de
                Ilojkic@veinst.hr
                Thomas.Mueller@fli.de
                Journal
                BMC Vet Res
                BMC Vet. Res
                BMC Veterinary Research
                BioMed Central (London )
                1746-6148
                13 March 2018
                13 March 2018
                2018
                : 14
                : 90
                Affiliations
                [1 ]IDT Biologika GmbH, Am Pharmapark, 06861 Dessau Rosslau, Germany
                [2 ]GRID grid.417834.d, Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institute, ; Südufer 10, 17493 Greifswald, Insel Riems Germany
                [3 ]ISNI 0000 0004 0367 0309, GRID grid.417625.3, Department of Virology, Croatian Veterinary Institute, ; Svaská cesta 143, 10000 Zagreb, Croatia
                Author information
                http://orcid.org/0000-0002-9662-3127
                Article
                1417
                10.1186/s12917-018-1417-0
                5850969
                29534727
                b122c40f-7c7c-4dda-aab9-0709395ba387
                © The Author(s). 2018

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 18 January 2018
                : 8 March 2018
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2018

                Veterinary medicine
                rabies,mongoose,oral vaccination,safety,spbn gasgas
                Veterinary medicine
                rabies, mongoose, oral vaccination, safety, spbn gasgas

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