title: Linear PADRE T helper epitope and carbohydrate B cell epitope conjugates induce specific high titer IgG antibody responses.
abstract: Linear carbohydrate-peptide constructs based on the 13 amino acid nonnatural pan DR epitope (PADRE) and carbohydrate B cell epitopes are demonstrated to be potent immunogens. These data support our belief that PADRE should be considered as an alternative to more complex carriers for use in prophylaxis and therapeutic vaccines. Two model carbohydrate-PADRE glycoconjugates were used to demonstrate that PADRE could effectively provide T cell help for carbohydrate-specific Ab responses. Conjugates of PADRE covalently linked to the human milk oligosaccharide, lacto-N-fucopentose II or a dodecasaccharide derived from Salmonella typhimurium O-Ag induced high titer IgG Ab responses in mice, which were comparable to glycoconjugates employing human serum albumin (HSA) as the carrier protein. Different adjuvants, in combination with PADRE conjugates, allowed for the modulation of the isotype profile with alum supporting an IgG1 profile; QS-21 an IgG2a, 2b profile, while an alum/QS-21 mixture generated a balanced IgG1/IgG2b isotype profile. As defined by binding to synthetic glycoconjugates, dodecasaccharide-specific Abs exhibited fine specificity similar to protective polyclonal Ab responses previously reported for dodecasaccharide-protein conjugates. The same Abs bound to intact S. typhimurium cells, suggesting that biologically relevant specificities were produced. The affinity of the dodecasaccharide-specific Abs was further shown to be comparable to that of a well-characterized, high affinity monoclonal anti-carbohydrate Ab recognizing the same epitope.
authors.affiliation: Epimmune, Inc., San Diego, CA 92121, USA. jalexander@epimmune.com
authors.forename: J
authors.initials: J
authors.lastname: Alexander
authors.forename: M F
authors.initials: MF
authors.lastname: del Guercio
authors.forename: A
authors.initials: A
authors.lastname: Maewal
authors.forename: L
authors.initials: L
authors.lastname: Qiao
authors.forename: J
authors.initials: J
authors.lastname: Fikes
authors.forename: R W
authors.initials: RW
authors.lastname: Chesnut
authors.forename: J
authors.initials: J
authors.lastname: Paulson
authors.forename: D R
authors.initials: DR
authors.lastname: Bundle
authors.forename: S
authors.initials: S
authors.lastname: DeFrees
authors.forename: A
authors.initials: A
authors.lastname: Sette
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002240:Carbohydrate Sequence; D002352:Carrier Proteins / Q000276:immunology; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D006001:Glycoconjugates / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis*; D007132:Immunoglobulin Isotypes / Q000096:biosynthesis; D017780:Malaria Vaccines / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000276:immunology; D012709:Serum Albumin / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology* / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D002352:Carrier Proteins; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D006001:Glycoconjugates; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D017780:Malaria Vaccines; D009844:Oligosaccharides; C401223:PADRE 45; D012709:Serum Albumin; C041762:lacto-N-fucopentaose II