title: Exploitation of bacterial N-linked glycosylation to develop a novel recombinant glycoconjugate vaccine against Francisella tularensis.
abstract: Glycoconjugate-based vaccines have proved to be effective at producing long-lasting protection against numerous pathogens. Here, we describe the application of bacterial protein glycan coupling technology (PGCT) to generate a novel recombinant glycoconjugate vaccine. We demonstrate the conjugation of the Francisella tularensis O-antigen to the Pseudomonas aeruginosa carrier protein exotoxin A using the Campylobacter jejuni PglB oligosaccharyltransferase. The resultant recombinant F. tularensis glycoconjugate vaccine is expressed in Escherichia coli where yields of 3 mg l(-1) of culture were routinely produced in a single-step purification process. Vaccination of BALB/c mice with the purified glycoconjugate boosted IgG levels and significantly increased the time to death upon subsequent challenge with F. tularensis subsp. holarctica. PGCT allows different polysaccharide and protein combinations to be produced recombinantly and could be easily applicable for the production of diverse glycoconjugate vaccines.
authors.affiliation: Department of Pathogen Molecular Biology, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
authors.forename: Jon
authors.initials: J
authors.lastname: Cuccui
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Thomas
authors.forename: Madeleine G
authors.initials: MG
authors.lastname: Moule
authors.forename: Riccardo V
authors.initials: RV
authors.lastname: D'Elia
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Laws
authors.forename: Dominic C
authors.initials: DC
authors.lastname: Mills
authors.forename: Diane
authors.initials: D
authors.lastname: Williamson
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Atkins
authors.forename: Joann L
authors.initials: JL
authors.lastname: Prior
authors.forename: Brendan W
authors.initials: BW
authors.lastname: Wren
mesh_terms: D036002:ADP Ribose Transferases / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001427:Bacterial Toxins / Q000276:immunology* / Q000378:metabolism; D001428:Bacterial Vaccines / Q000737:chemistry / Q000276:immunology; D016123:Campylobacter jejuni / Q000201:enzymology; D004926:Escherichia coli / Q000378:metabolism; D005098:Exotoxins / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005604:Francisella tularensis / Q000276:immunology* / Q000378:metabolism; D006031:Glycosylation; D006602:Hexosyltransferases / Q000378:metabolism; D007074:Immunoglobulin G / Q000097:blood; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D019081:O Antigens / Q000276:immunology* / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000276:immunology / Q000378:metabolism; D013678:Technology, Pharmaceutical; D014406:Tularemia / Q000276:immunology / Q000517:prevention & control*; D018074:Vaccines, Conjugate / Q000737:chemistry / Q000276:immunology; D037521:Virulence Factors / Q000276:immunology* / Q000378:metabolism; D000097668:Pseudomonas aeruginosa Exotoxin A
chemical_list: D036002:ADP Ribose Transferases; D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D005098:Exotoxins; D006602:Hexosyltransferases; D007074:Immunoglobulin G; D008565:Membrane Proteins; D019081:O Antigens; D018074:Vaccines, Conjugate; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase
keywords: Francisella tularensis; glycoconjugate; protein glycan coupling technology; vaccine