BACKGROUND Stx toxin is a member of the AB5 family of bacterial toxins: the active A subunit has N-glycosidase activity against 28S rRNA, resulting in inhibition of protein synthesis in eukaryotic cells, and the pentamer ligand B subunits (StxB) bind to globotria(tetra)osylceramide receptors (Gb3/Gb4) on the cell membrane. Shiga toxin-producing Escherichia coli strains (STEC) may produce Stx1 a...

The Rockefeller University Press $30.00 J. Cell Biol. Vol. 209 No. 4 563–577 www.jcb.org/cgi/doi/10.1083/jcb.201410026 JCB 563 *M.K. Dennis and A.R. Mantegazza contributed equally to this paper. **M.S. Marks and S.R.G. Setty contributed equally to this paper. Correspondence to Michael S. Marks: [email protected] Abbreviations used in this paper: AF488, Alexa Fluor 488; AP-3, adaptor pro...

Shigella cytotoxin was purified from the culture medium of Shigella Shiga 60 R and from the bacterial pellet by an extensive purification scheme involving chromatography on Cibachron Blue F3GA, chitin, and DEAE-cellulose columns, sucrose gradient centrifugation, and finally polyacrylamide gel electrophoresis under nondenaturing conditions. In sodium dodecyl sulfate polyacrylamide gels, purified...

Gut-derived lymphocytes transiently migrate through the peripheral circulation before homing back to mucosal sites and can be detected using an ELISPOT-based antibody secreting cell (ASC) assay. Alternatively, transiently circulating lymphocytes may be cultured in vitro, and culture supernatants may be assayed for antigen-specific responses (antibody in lymphocyte supernatant [ALS] assay). The ...

Retrograde transport is where proteins and lipids are transported back from the plasma membrane (PM) and endosomes to the Golgi, and crucial for a diverse range of cellular functions. Recycling endosomes (REs) serve as a sorting station for the retrograde transport and we recently identified evection-2, an RE protein with a pleckstrin homology (PH) domain, as an essential factor of this pathway...

Shigellosis is a severe diarrheal disease caused by members of the genus Shigella , with at least 80 million cases and 700,000 deaths annually around world. The type III secretion system (T3SS) primary virulence factor used shigellae, we have previously demonstrated that vaccination T3SS proteins IpaB IpaD, along an IpaD/IpaB fusion protein (DBF), protects mice from infection in lethal pulmonar...

Infection with Vibrio cholerae and oral cholera vaccines (OCVs) induce transient circulating plasmablast responses that peak within approximately 7 days after infection or vaccination. We previously demonstrated that plasmablast responses strongly correlate with subsequent levels of V. cholerae-specific duodenal antibodies up to 6 months after V. cholerae infection. Hence, plasmablast responses...

A chimeric protein consisting of a divalent pertussis toxin (PT) S1 fragment linked to the cholera toxin (Ctx) A(2)B fragment was constructed. The chimera induced a mucosal immunoglobulin A (IgA) and a serum IgG immune response to PT and CtxB in BALB/c mice following intranasal immunization. The immune sera neutralized PT in vitro. In the mouse model of Bordetella pertussis respiratory infectio...

We investigated whether eyedrop vaccination using modified outer membrane vesicles (mOMVs) is effective for protecting against hemolytic uremic syndrome (HUS) caused by enterohemorrhagic E. coli (EHEC) O157:H7 infection. Modified OMVs and waaJ-mOMVs were prepared from cultures of MsbB- and Shiga toxin A subunit (STxA)-deficient EHEC O157:H7 bacteria with or without an additional waaJ mutation. ...