@article {Baram816553, author = {Noam Baram and Hadar Cohen and Liat Edry-Botzer and Dor Salomon and Motti Gerlic}, title = {Vibrio pore-forming leukocidin activates pyroptotic cell death via the NLRP3 inflammasome}, elocation-id = {816553}, year = {2019}, doi = {10.1101/816553}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Cell death mechanisms are central to combat infectious microbes and to drive pathological inflammation. One such mechanism, the inflammasome, controls infection through either activation of caspase-1 and the subsequent secretion of the mature pro-inflammatory cytokine, interleukin 1β (IL-1β), or by stopping the dissemination of intracellular pathogens by inducing pyroptotic cell death in infected cells. Hemolysins, which are pore-forming toxins (PFTs), target the host cell plasma membrane by producing pores with different diameters. These pores alter the permeability of the target membrane, often leading to cell death. We previously discovered a functional and potent pore-forming, leukocidin domain-containing hemolysin produced by the Gram-negative marine bacterium Vibrio proteolyticus (V. proteolyticus), termed VPRH. Although leukocidin domains are found in other known PFTs, VPRH constitutes a distinct, understudied class within the leukocidin superfamily. Since PTFs of other pathogens were shown to induce cell death by activating the inflammasome pathway, we hypothesized that VPRH-induced cell death is mediated by direct activation of the inflammasome in mammalian immune host cells. Indeed, we found that VPRH induced a two-step cell death in primary macrophages. The first, a rapid step, was mediated by activating the NLRP3 inflammasome, leading to caspase-1 activation and GSDMD cleavage that resulted in IL-1β secretion and pyroptotic cell death. The second step was independent of the inflammasome; however, its mechanism remains unknown. This study sets the foundation for better understanding the immunological consequences of inflammasome activation by a new leukocidin class of toxins.}, URL = {https://www.biorxiv.org/content/early/2019/10/24/816553}, eprint = {https://www.biorxiv.org/content/early/2019/10/24/816553.full.pdf}, journal = {bioRxiv} }