Formulation and characterisation of Bordetella pertussis fimbriae as novel carrier proteins for Hib conjugate vaccines.
Haemophilus influenzae type b (Hib) capsular polysaccharide (polyribosylribitol phosphate, PRP) is the active component of conjugate vaccines that have proven successful in preventing invasive Hib disease. Conjugation of PRP to a protein carrier greatly improves its immunogenicity providing protection in infants and subsequent antibody maturation upon boosting. In this study, fimbriae isolated from Bordetella pertussis have been assessed as novel carrier proteins. These proteins are components of some acellular pertussis vaccines and clinical trials have indicated that fimbriae could be important protective antigens against whooping cough. Fimbriae (Fim2 and Fim3) purified from B. pertussis were dissociated in 6 M guanidine hydrochloride, pH 10.5, to produce proteins of defined size and to facilitate the production and characterisation of the conjugates. Both carbodiimide-mediated coupling and reductive amination were used to conjugate PRP to dissociated fimbriae. Efficiency of conjugation was determined by size exclusion chromatography followed by protein and polysaccharide analysis of fractionated components. Immunisation of rabbits with dissociated fimbriae-PRP conjugates (D.fim-PRP) produced high anti-fimbrial and anti-PRP IgG titres. Use of a D.fim-PRP conjugate could protect against Hib disease and may also augment protection against B. pertussis.
Crowley-Luke
A
A
Centre for Applied Microbiology and Research, SP4 0JG, Salisbury, UK. annette.crowley-luke@camr.org.uk
Reddin
K
K
Gorringe
A
A
Hudson
M J
MJ
Robinson
A
A
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000302:isolation & purification; D016667:Bacterial Capsules; D001886:Bordetella pertussis / Q000276:immunology*; D002352:Carrier Proteins / Q000008:administration & dosage / Q000302:isolation & purification; D005260:Female; D010861:Fimbriae, Bacterial / Q000276:immunology*; D006192:Haemophilus Infections / Q000276:immunology / Q000517:prevention & control; D018073:Haemophilus Vaccines / Q000008:administration & dosage*; D019741:Haemophilus influenzae type b / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007231:Infant, Newborn; D011134:Polysaccharides / Q000008:administration & dosage / Q000276:immunology / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000008:administration & dosage*; D011817:Rabbits; D018074:Vaccines, Conjugate / Q000008:administration & dosage; D014917:Whooping Cough / Q000276:immunology / Q000517:prevention & control
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D002352:Carrier Proteins; D018073:Haemophilus Vaccines; C044237:Haemophilus influenzae type b polysaccharide vaccine; D007074:Immunoglobulin G; D011134:Polysaccharides; D011135:Polysaccharides, Bacterial; D018074:Vaccines, Conjugate; C008984:polyribitol phosphate