title: A putative amino acid ABC transporter substrate-binding protein, NMB1612, from Neisseria meningitidis, induces murine bactericidal antibodies against meningococci expressing heterologous NMB1612 proteins.
abstract: The nmb1612 (NEIS1533) gene encoding the ~27-kDa putative amino acid ATP-binding cassette (ABC) transporter, periplasmic substrate-binding protein from Neisseria meningitidis serogroup B (MenB) strain MC58 was cloned and expressed in Escherichia coli, and the purified recombinant (r)NMB1612 was used for animal immunization studies. Immunization of mice with rNMB1612 adsorbed to Al(OH)3 and in liposomes with and without MPLA, induced antiserum with bactericidal activity in an assay using baby rabbit complement, against the homologous strain MC58 (encoding protein representative of Allele 62) and killed heterologous strains encoding proteins of three other alleles (representative of Alleles 1, 64 and 68), with similar SBA titres. However, strain MC58 was not killed (titre <4) in a human serum bactericidal assay (hSBA) using anti-rNMB1612 sera, although another strain (MC168) expressing the same protein was killed (median titres of 16-64 in the hSBA). Analysis of the NMB1612 amino acid sequences from 4351 meningococcal strains in the pubmlst.org/Neisseria database and a collection of 13 isolates from colonized individuals and from patients, showed that antibodies raised against rNMB1612 could potentially kill at least 72% of the MenB strains in the complete sequence database. For MenB disease occurring specifically in the UK from 2013 to 2015, >91% of the isolates causing disease in this recent period expressed NMB1612 protein encoded by Allele 1 and could be potentially killed by sera raised to the recombinant antigen in the current study. The NMB1612 protein was surface-accessible and expressed by different meningococcal strains. In summary, the properties of (i) NMB1612 protein conservation and expression, (ii) limited amino acid sequence variation between proteins encoded by different alleles, and (iii) the ability of a recombinant protein to induce cross-strain bactericidal antibodies, would all suggest a promising antigen for consideration for inclusion in new meningococcal vaccines.
authors.affiliation: Neisseria Research, Molecular Microbiology, Division of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton Medical School, Southampton SO166YD, United Kingdom.
authors.forename: Miao-Chiu
authors.initials: MC
authors.lastname: Hung
authors.affiliation: Neisseria Research, Molecular Microbiology, Division of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton Medical School, Southampton SO166YD, United Kingdom.
authors.forename: María Victoria
authors.initials: MV
authors.lastname: Humbert
authors.affiliation: Neisseria Research, Molecular Microbiology, Division of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton Medical School, Southampton SO166YD, United Kingdom.
authors.forename: Jay R
authors.initials: JR
authors.lastname: Laver
authors.affiliation: Neisseria Research, Molecular Microbiology, Division of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton Medical School, Southampton SO166YD, United Kingdom.
authors.forename: Renee
authors.initials: R
authors.lastname: Phillips
authors.affiliation: Neisseria Research, Molecular Microbiology, Division of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton Medical School, Southampton SO166YD, United Kingdom.
authors.forename: John E
authors.initials: JE
authors.lastname: Heckels
authors.affiliation: Neisseria Research, Molecular Microbiology, Division of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton Medical School, Southampton SO166YD, United Kingdom. Electronic address: mc4@soton.ac.uk.
authors.forename: Myron
authors.initials: M
authors.lastname: Christodoulides
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000276:immunology*; D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000536:Aluminum Hydroxide / Q000008:administration & dosage; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D001770:Blood Bactericidal Activity; D003001:Cloning, Molecular; D003429:Cross Reactions; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D064789:Immunity, Heterologous; D008081:Liposomes / Q000008:administration & dosage; D022401:Meningococcal Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D008807:Mice, Inbred BALB C; D038541:Neisseria meningitidis, Serogroup B / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
chemical_list: D018528:ATP-Binding Cassette Transporters; D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D008081:Liposomes; D022401:Meningococcal Vaccines; D014614:Vaccines, Synthetic; D000536:Aluminum Hydroxide
keywords: Amino acid ABC transporter; Bactericidal activity; Meningococcal vaccine; Neisseria meningitidis; Sequence diversity; Substrate-binding protein