title: Treatment with succinic anhydride improves the immunogenicity of Shigella flexneri type 2a O-specific polysaccharide-protein conjugates in mice.
abstract: Seroepidemiological data and a clinical trial with a Shigella sonnei O-specific polysaccharide (O-SP)-Pseudomonas aeruginosa recombinant exoprotein A (rEPA) conjugate provide evidence that a critical level of immunoglobulin G (IgG) lipopolysaccharide (LPS) antibodies in serum confers protection against shigellosis. We evaluated the immunogenicity of conjugates whose carrier proteins and O-SPs were treated with succinic anhydride (SA), which reacts with amino groups at neutral pH to form amide-linked carboxyls (succinylation). Conjugates were synthesized with either of two genetically inactivated medically useful toxins, the diphtheria protein CRM9 or rEPA, bound to the O-SP of Shigella flexneri type 2a. Conjugates composed of the succinylated protein, succinylated O-SP, or both succinylated components were administered to mice by a clinically relevant scheme, and their levels of serum IgG anti-LPS and anti-proteins were assayed 7 days after the second and third injections. CRM9 served as a more immunogenic carrier than rEPA. Conjugates composed of succinylated components were more immunogenic than the conjugates composed of the native components. SA treatment of both the carrier protein and the O-SP did not confer an advantage over the succinylated protein alone. Conjugates prepared with native proteins, in general, elicited slightly higher levels of IgG protein antibodies than conjugates composed of the SA-treated proteins.
authors.affiliation: National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892-2720, USA.
authors.forename: D
authors.initials: D
authors.lastname: Pavliakova
authors.forename: C
authors.initials: C
authors.lastname: Chu
authors.forename: S
authors.initials: S
authors.lastname: Bystricky
authors.forename: N W
authors.initials: NW
authors.lastname: Tolson
authors.forename: J
authors.initials: J
authors.lastname: Shiloach
authors.forename: J B
authors.initials: JB
authors.lastname: Kaufman
authors.forename: D A
authors.initials: DA
authors.lastname: Bryla
authors.forename: J B
authors.initials: JB
authors.lastname: Robbins
authors.forename: R
authors.initials: R
authors.lastname: Schneerson
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001428:Bacterial Vaccines / Q000276:immunology*; D005260:Female; D051379:Mice; D019081:O Antigens / Q000276:immunology*; D012763:Shigella flexneri / Q000276:immunology*; D013387:Succinic Anhydrides / Q000494:pharmacology*; D018074:Vaccines, Conjugate / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D019081:O Antigens; D013387:Succinic Anhydrides; D018074:Vaccines, Conjugate; C031801:succinic anhydride