title: Salmonella enterica serovar enteritidis core O polysaccharide conjugated to H:g,m flagellin as a candidate vaccine for protection against invasive infection with S. enteritidis.
abstract: Nontyphoidal Salmonella enterica serovars Enteritidis and Typhimurium are a common cause of gastroenteritis but also cause invasive infections and enteric fever in certain hosts (young children in sub-Saharan Africa, the elderly, and immunocompromised individuals). Salmonella O polysaccharides (OPS) and flagellar proteins are virulence factors and protective antigens. The surface polysaccharides of Salmonella are poorly immunogenic and do not confer immunologic memory, limitations overcome by covalently attaching them to carrier proteins. We conjugated core polysaccharide-OPS (COPS) of Salmonella Enteritidis lipopolysaccharide (LPS) to flagellin protein from the homologous strain. COPS and flagellin were purified from a genetically attenuated (ΔguaBA) "reagent strain" (derived from an isolate from a patient with clinical bacteremia) engineered for increased flagellin production (ΔclpPX). Conjugates were constructed by linking flagellin monomers or polymers at random COPS hydroxyls with various polysaccharide/protein ratios by 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) or at the 3-deoxy-d-manno-octulosonic acid (KDO) terminus by thioether chemistry. Mice immunized on days 0, 28, and 56 with COPS-flagellin conjugates mounted higher anti-LPS IgG levels than mice receiving unconjugated COPS and exhibited high antiflagellin IgG; anti-LPS and antiflagellin IgG levels increased following booster doses. Antibodies generated by COPS-flagellin conjugates mediated opsonophagocytosis of S. Enteritidis cells into mouse macrophages. Mice immunized with flagellin alone, COPS-CRM₁₉₇, or COPS-flagellin conjugates were significantly protected from lethal challenge with wild-type S. Enteritidis (80 to 100% vaccine efficacy).
authors.affiliation: Center for Vaccine Development, University of Maryland, Baltimore, Baltimore, MD 21201, USA. rsimon@medicine.umaryland.edu
authors.forename: Raphael
authors.initials: R
authors.lastname: Simon
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Tennant
authors.forename: Jin Y
authors.initials: JY
authors.lastname: Wang
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Schmidlein
authors.forename: Andrew
authors.initials: A
authors.lastname: Lees
authors.forename: Robert K
authors.initials: RK
authors.lastname: Ernst
authors.forename: Marcela F
authors.initials: MF
authors.lastname: Pasetti
authors.forename: James E
authors.initials: JE
authors.lastname: Galen
authors.forename: Myron M
authors.initials: MM
authors.lastname: Levine
mesh_terms: D000818:Animals; D021461:Animals, Outbred Strains; D000907:Antibodies, Bacterial / Q000097:blood*; D002460:Cell Line; D005260:Female; D005408:Flagellin / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007114:Immunization; D018796:Immunoconjugates / Q000008:administration & dosage* / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D019081:O Antigens / Q000737:chemistry / Q000276:immunology*; D009895:Opsonin Proteins / Q000378:metabolism; D010587:Phagocytosis; D012481:Salmonella Infections, Animal / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D022562:Salmonella Vaccines / Q000008:administration & dosage* / Q000276:immunology; D012477:Salmonella enteritidis / Q000276:immunology* / Q000472:pathogenicity
chemical_list: D000907:Antibodies, Bacterial; D018796:Immunoconjugates; D007074:Immunoglobulin G; D019081:O Antigens; C062656:O-specific polysaccharide, Salmonella; D009895:Opsonin Proteins; D022562:Salmonella Vaccines; D005408:Flagellin