Ic cells. Purification through a 12 step sucrose gradient was performed prior to conditioning in vitro and in vivo.Introduction: Infections by two Gram-negative intracellular bacterial pathogens B7-H2/ICOSLG Proteins manufacturer Piscirickettsia salmonis and Francisella noatunensis, are causing significant complications in aquaculture world-wide. F. noatunensis sp hampers the development of fish farming based on cod in and is deleterious to tilapia. P. salmonis infections have been devastating for salmon aquaculture. As of today no productive treatment options are out there against the illnesses. Each P. salmonis and F. noatunensis secrete membrane vesicles (MV). Bacterial MV has been reported as possible vaccine candidates for any range of host including humans, mice and fish against infection triggered by intracellular pathogenic bacteria as they induce both a humoral and cellular immunity.ISEV2019 ABSTRACT BOOKMethods: We’ve got isolated MVs from each Francisella and Piscirickettsia by the ultracentrifugation Technique. The MVs have been characterized by their size distribution, by transmission electron microscopy (TEM) and proteomics. Their toxicity have been tested by injecting MVs into both our zebrafish vaccine and challenge model at the same time as in cod, tilapia and salmon. A vaccine trail was performed initially in our zebrafish model, and then in cod, tilapia and salmon. Outcomes: 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 identified in the population as investigated by TEM. Additional, immune-gold labelling revealed the presence of DNA in each populations. Proteomics analysis revealed that the MV content material varied between bacterial strains. Immunization with MV gave protection against illness triggered by both P. salmonis and F. noatunensis in our zebrafish model, having said that, did not guard cod, tilapia nor salmon. Summary/Conclusion: The MVs from P. salmonis and F. noatunensis revealed a comparable size distribution and that the content includes various bacterial virulence variables also as DNA that can be transferred towards the host. As for their immunogenic properties this appears to vary CD74 Proteins custom synthesis amongst the vaccine and challenge model in comparison to the all-natural hosts. The use of the MVs as vaccines in their organic hosts including strain-specificity and cross-immunity need to have further 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 Health-related Analysis, 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 regular epithelium. For this, colonic epithelial cells in the T84 line had been grown on Transwell filters to create transepithelial electrical resistance (TEER), a measure of epithelial monolayer integrity. The cells were then cocultured with Alexa Fluor-labelled OMVs from the gastric pathogen, Helicobacter pylori. Final results: We showed that H. pylori OMVs readily entered polarised epithelial cells, but had no impact around the TEER nor permeability.