Abstract
This thematic Project includes 29 subprojects, focusing on emerging zoonotic viruses of the family Bunyaviridae, distributed in 3 genera: Orthobunyavirus, Phlebovirus and Hantavirus. The project involves 3 Principal Researchers, 55 Brazilian and international collaborators, and will take 4 years. For studies on the genus Orthobunyavirus, serological diagnosis of Oropouche virus (OROV) by ELISA will be developed using a recombinant N protein of OROV as antigen. This will enable sero-epidemiological surveys in patients, horses, small mammals and birds. The recombinant N protein of OROV will also be tested as an immunogen in mice, and the mechanism of its apoptotic effect will be studied in vitro. The antiviral activity of 10,000 compounds for Orthobunyavirus will be tested. Cell biology in vitro studies will be done to verify sites of OROV assembly and budding in host cells, and to identify factors that regulate intracellular traffic of proteins involved in those processes. A fluorescent OROV will also be produced by reverse genetics for in vitro and in vivo studies. Molecular epidemiology and phylogenetic studies of orthobunyaviruses based on complete genomes will be done by next-generation sequencing. A real time RT-PCR assay for detection of Brazilian orthobunyaviruses of serogroups C and Simbu will also be developed. Studies on the genus Phlebovirus will include molecular epidemiology and phylogeny, development of a real time RT-PCR to assess their occurrence in clinical samples from patients with acute febrile illness, testing of the antiviral activity of of 10,000 compounds and development of an experimental mouse model of infection by Candiru virus. Studies will also be done to detect assembly/budding sites of Candiru virus. Studies on the genus Hantavirus will include virus detection in hantavirus cardio-pulmonary syndrome (HCPS) and study of its pathophysiology based on levels of cytokines in patient blood samples. Importantly, we plan to characterize genetically and morphologically the Araraquara (ARQV) strain of antavirus and evaluate the immunogenicity of a recombinant N protein of ARQV in mice. We also will test the antiviral activity of 10,000 compounds against The strain Rio Mamore of hantavirus). Field capture of small mammals and their ectoparasites will be done to investigate Hantavirus infection, especially in Necromys lasiurus. A molecular epidemiology study will be done in cooperation with scientists from other national and international (USA, Finland and Colombia) institutions, based on computational analysis of the evolution and geographical distribution of Brazilian hantaviruses. Cell biology studies will hopefully reveal details of Hantavirus assembly and budding. (AU)
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