R and temporal disturbances to the monolayer’s integrity within thirty min post infection. No disturbances have been seen upon addition of non-infected EVs. Summary/conclusion: Our study demonstrates that EVs-derived from ZIKV-infected cells are able to transfer proteins and viral RNA to recipient cells. Considering that the two IEVs and viral particles can induce similar changes on barrier’s integrity it’s doable that IEVs are concerned in an option mechanism of ZIKV transmission.PS02.09= OWP2.Deciphering the function of extracellular vesicles over the blood rain barrier all through Zika virus infection Antonios Fikatas, Sam Noppen, Peter Vervaeke, Jordi Doijen, Mohammed Benkheil, Christophe Pannecouque and Dominique Schols Laboratory of Virology and Chemotherapy, Rega Institute, KU Leuven, Belgium, Leuven, BelgiumPS02.10=OWP2.In vivo testing of OMV-based vaccine prototypes against Gallibacterium anatis Fabio Antenuccia, Homa Arakb, Jianyang Gaob, Toloe Allahghadryb, Ida Th nerb and Anders Miki BojesencaUniversity of IDO Proteins Storage & Stability Copenhagen, K enhavn S, Denmark; bUniversity of Copenhagen, Copenhagen, Denmark; cUniversity of Copenhagen, Copenhagen, USAIntroduction: The association of Zika virus (ZIKV) with extreme neurological problems has acquired greater curiosity in excess of the final decade. Even so, the mechanism by which ZIKV crosses the blood rain barrier (BBB) and reaches the brain stays to get elucidated. It’s acknowledged that viruses include viral material in extracellular vesicles (EVs) being a spreading system. These membrane-enclosed vesicles perform a important position in intercellular communication. Presently, there is a lack of awareness to the possible involvement of EVs in ZIKV pathogenesis. Our review aims to unravel the position of EVs in ZIKV RNA transmission to the brain, via the BBB. Solutions: Human brain microvascular endothelial cells (HBMEC/D3) have been utilized in our examine since they represent the BBB in vitro. 3 different EV isolation strategies (precipitation kit, density gradient and size exclusion chromatography mixed together with the density gradient) were carried out. Western blot, Transmission electron microscopy and Nanosight monitoring examination confirmed the presence of EVs in the supernatant of HBMEC/D3 cells. The presence of ZIKV RNA in infected-EVs (IEVs) was evaluated by immunofluorescence and qPCR. In addition, the impact of IEVs on the BBB was assessed using a label-free impedance-based biosensor (ECIS, Applied BioPhysics). Final results: We confirmed the presence of viral GP-Ib alpha/CD42b Proteins Recombinant Proteins elements in our IEVs, together with the NS1 and E proteins of ZIKV. The obtained IEVs have been in a position to re-infectIntroduction: Outer membrane vesicles (OMVs) are made from the majority of Gram-negative bacteria. Because of the antigenic similarity in between OMVs along with the bacterial outer membrane, OMVs have proven to become promising to the growth of novel vaccines towards bacterial pathogens. In this work, we describe the testing of OMV-based vaccine prototypes against Gallibacterium anatis, a Gram-negative pathogen of great veterinary interest. Solutions: OMVs have been isolated from a G. anatis hypervesiculating mutant utilizing a modified edition of the Hydrostatic Filtration protocol described by Musante et al. (2014). 120 16-week-old Lohmann-Brown chickens have been divided in six groups and immunized twice intramuscularly with different combinations of buffer (controls), OMVs and selected recombinant immunogens. Two weeks after second immunization, the effectiveness of the immunization regimes adopted was examined by difficult t.