title: Physico-chemical characterisation and immunogenicity of a multi-valent candidate vaccine against non-typeable Haemophilus influenzae and Moraxella catarrhalis.
abstract: The physico-chemical characteristics and immunogenicity of a candidate vaccine against otitis media, prepared from recombinant lipidated outer membrane proteins (rLP4 and rLP6) from non-typeable Haemophilus influenzae (NTHi) and of the ubiquitous cell surface protein UspA2 from Moraxella catarrhalis, were evaluated. Optical spectroscopy, size exclusion chromatography and gel electrophoresis were used to characterise the purified protein components and assess their purity and molecular sizes. The results showed that the three proteins were highly purified. Possible dimers in rLP4, dimers and multimers in rLP6 and UspA2 were detected. Small amounts of rLP4 and rLP6 dimers and most of UspA2 complexes remained tightly bound even after SDS treatment under reducing conditions. Immunogenicity studies showed that all proteins induced substantial antibody responses in mice immunised with AlPO4-adsorbed rLP4, rLP6 or UspA2 or a combination of these proteins. However, combination of these proteins resulted in a reduced response to rLP4 and rLP6, but not to UspA2, suggesting interference between these proteins which should be taken into consideration during the development and evaluation of this vaccine.
authors.affiliation: Division of Bacteriology, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. fmawas@nibsc.ac.uk
authors.forename: Fatme
authors.initials: F
authors.lastname: Mawas
authors.forename: Mei Mei
authors.initials: MM
authors.lastname: Ho
authors.forename: Rachel
authors.initials: R
authors.lastname: Huskisson
authors.forename: Manolya
authors.initials: M
authors.lastname: Saydam
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Corbel
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D017607:Aluminum Compounds / Q000494:pharmacology; D000818:Animals; D001428:Bacterial Vaccines / Q000737:chemistry* / Q000276:immunology*; D015153:Blotting, Western; D049109:Cell Proliferation; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002850:Chromatography, Gel; D002942:Circular Dichroism; D016207:Cytokines / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D018073:Haemophilus Vaccines / Q000737:chemistry* / Q000276:immunology*; D007111:Immunity, Cellular / Q000187:drug effects; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D001936:Moraxella catarrhalis / Q000276:immunology*; D010710:Phosphates / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D017778:Vaccines, Combined / Q000737:chemistry / Q000276:immunology; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D017607:Aluminum Compounds; D001428:Bacterial Vaccines; D016207:Cytokines; D018073:Haemophilus Vaccines; D007074:Immunoglobulin G; D008565:Membrane Proteins; D010710:Phosphates; D017778:Vaccines, Combined; D014614:Vaccines, Synthetic; C012714:aluminum phosphate