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Multiple studies have established the importance of the tol‐pal gene cluster in bacterial cell membrane integrity and outer membrane vesicle (OMV) formation in Escherichia coli. In contrast, the functions of Tol‐Pal proteins in pathogenic organisms, including those of the Epsilonproteobacteria, remain poorly if at all defined. The aim of this study was to characterize the roles of two key components of the Tol‐Pal system, TolB and Pal, in OMV formation in the pathogenic bacterium, Helicobacter pylori.

H. pylori ΔtolB, Δpal and ΔtolBpal mutants, as well as complemented strains, were generated and assessed for changes in morphology and OMV production by scanning electron microscopy and enzyme‐linked immunoassay (ELISA), respectively. The protein content and pro‐inflammatory properties of OMVs were determined by mass spectroscopy and interleukin‐8 (IL‐8) ELISA on culture …