Extraintestinal pathogenic E. coli (ExPEC) cause systemic disease among birds, humans, and mammals, including urinary tract infection, septicemia, neonatal meningitis in human, aerosacculitis and polyserositis in chicken. Conjugative ColV-related plasmids are responsible for ExPEC virulence. ColV-related virulence factors enhance ExPECs colonization and fitness during the infection. The hlyF gene in ColV-related plasmids is an epidemiology marker for highly virulent ExPEC responsible for neonatal meningitis in humans and colibacillosis in chicken. This plasmid carrying hlyF is also present in the enterohemorrhagic E. coli (EHEC) of serotype O80:H2 which has recently emerged in France and represents a new threat in terms of public health. We have shown that HlyF is a fake hemolysin but a real virulence factor that can directly mediate the production of outer membrane vesicles (OMVs). These OMVs induce a massive autophagy blockade before the fusion of the autophagosome. In parallel, OMVs treatments induce the production of pro-inflammatory cytokines and chemokines by a massive activation of the non-canonical inflammasome pathway. Both phenotypes require the catalytic activity of HlyF since they are not observed with OMVs from catalytically inactive hlyF+ E. coli. While the autophagosome and the inflammasome implement checks and balances on each other, these specific OMVs alter these checks and tip the balance toward a massive activation of the NLRP3-inflammasome. Comprehensive studies are still required to tease out the relationships between autophagy and inflammasomes, and how these specific OMVs may facilitate toxins and bacterial dissemination across host barriers.