Ic cells. Purification via a 12 step sucrose gradient was performed prior to conditioning in vitro and in vivo.Introduction: Infections by two Gram-negative intracellular bacterial mGluR8 MedChemExpress pathogens Piscirickettsia salmonis and Francisella noatunensis, are causing major complications in aquaculture world-wide. F. noatunensis sp hampers the development of fish farming depending on cod in and is deleterious to tilapia. P. salmonis infections have been devastating for salmon aquaculture. As of currently no effective treatments are accessible against the diseases. Each P. salmonis and F. noatunensis secrete membrane vesicles (MV). Bacterial MV has been reported as potential vaccine candidates for any selection of host such as humans, mice and fish against infection caused by intracellular pathogenic bacteria as they induce each a humoral and cellular immunity.ISEV2019 ABSTRACT BOOKMethods: We have isolated MVs from both Francisella and Piscirickettsia by the ultracentrifugation Approach. The MVs had been characterized by their size distribution, by transmission electron microscopy (TEM) and proteomics. Their toxicity were tested by injecting MVs into both our zebrafish vaccine and challenge model as well as in cod, tilapia and salmon. A vaccine trail was performed 1st in our zebrafish model, and after that in cod, tilapia and salmon. Final results: The MV size evaluation showed that the MVs size distribution ranged from 2050 nm in size with most ranging from 7000 nm. Both single and double membrane MV have been found inside the population as investigated by TEM. Further, immune-gold labelling revealed the presence of DNA in each populations. Proteomics analysis revealed that the MV content varied between bacterial strains. Immunization with MV gave protection against disease brought on by each P. salmonis and F. noatunensis in our zebrafish model, nevertheless, didn’t guard cod, tilapia nor salmon. Summary/Conclusion: The MVs from P. salmonis and F. noatunensis revealed a related size distribution and that the content contains different bacterial virulence components at the same time as DNA that may be transferred to the host. As for their immunogenic properties this seems to differ involving the vaccine and challenge model in comparison to the organic hosts. The use of the MVs as vaccines in their organic hosts for instance strain-specificity and cross-immunity want additional investigation. Funding: Analysis Council of Norway (RCN) and University of Oslo.OF14.Bacterial membrane vesicles enter polarised epithelial cells and provide their protein cargo to exosomes Lorinda Turnera, Nestor Solisb, Georg Rammc, Viola Oorschotc, Amanda De Paolia, Hassan Chaudhrya, Stuart Manneringd, Stuart Cordwellb, Maria Kaparakis-Liaskose and Richard Ferreroaa Hudson Institute of Healthcare Study, Melbourne, Australia; bThe University of Sydney, Sydney, Australia; cMonash University, Melbourne, Australia; dSt. Vincent’s Institute of Medical Study, Melbourne, Australia; 5Department of Physiology, Anatomy and Microbiology, La Trobe University, Melbourne, Australiaresistance and apical-basolateral polarity of standard RSK2 Purity & Documentation epithelium. For this, colonic epithelial cells of the T84 line were grown on Transwell filters to produce transepithelial electrical resistance (TEER), a measure of epithelial monolayer integrity. The cells had been then cocultured with Alexa Fluor-labelled OMVs in the gastric pathogen, Helicobacter pylori. Outcomes: We showed that H. pylori OMVs readily entered polarised epithelial cells, but had no impact on the TEER nor permeability.