title: Investigation into the Antigenic Properties and Contributions to Growth in Blood of the Meningococcal Haemoglobin Receptors, HpuAB and HmbR.
abstract: Acquisition of iron from host complexes is mediated by four surface-located receptors of Neisseria meningitidis. The HmbR protein and heterodimeric HpuAB complex bind to haemoglobin whilst TbpBA and LbpBA bind iron-loaded transferrin and lactoferrin complexes, respectively. The haemoglobin receptors are unevenly distributed; disease-causing meningococcal isolates encode HmbR or both receptors while strains with only HpuAB are rarely-associated with disease. Both these receptors are subject to phase variation and 70-90% of disease isolates have one or both of these receptors in an ON expression state. The surface-expression, ubiquity and association with disease indicate that these receptors could be potential virulence factors and vaccine targets. To test for a requirement during disease, an hmbR deletion mutant was constructed in a strain (MC58) lacking HpuAB and in both a wild-type and TbpBA deletion background. The hmbR mutant exhibited an identical growth pattern to wild-type in whole blood from healthy human donors whereas growth of the tbpBA mutant was impaired. These results suggest that transferrin is the major source of iron for N. meningitidis during replication in healthy human blood. To examine immune responses, polyclonal antisera were raised against His-tagged purified-recombinant variants of HmbR, HpuA and HpuB in mice using monolipopolysaccharide as an adjuvant. Additionally, monoclonal antibodies were raised against outer membrane loops of HmbR presented on the surface of EspA, an E. coli fimbrial protein. All antisera exhibited specific reactivity in Western blots but HmbR and HpuA polyclonal sera were reactive against intact meningococcal cells. None of the sera exhibited bactericidal activity against iron-induced wild-type meningococci. These findings suggest that the HmbR protein is not required during the early stages of disease and that immune responses against these receptors may not be protective.
authors.affiliation: Department of Genetics, University of Leicester, Leicester, United Kingdom.
authors.forename: Fadil A
authors.initials: FA
authors.lastname: Bidmos
authors.affiliation: National Institute for Biological Standards and Control, Potters Bar, United Kingdom.
authors.forename: Hannah
authors.initials: H
authors.lastname: Chan
authors.affiliation: Department of Genetics, University of Leicester, Leicester, United Kingdom.
authors.forename: Uta
authors.initials: U
authors.lastname: Praekelt
authors.affiliation: Department of Genetics, University of Leicester, Leicester, United Kingdom.
authors.forename: Isfahan
authors.initials: I
authors.lastname: Tauseef
authors.affiliation: Department of Infection, Inflammation and Immunity, University of Leicester, Leicester, United Kingdom; Microbiology Department, Faculty of Pharmacy, Mansoura University, Mansoura, Egypt.
authors.forename: Youssif M
authors.initials: YM
authors.lastname: Ali
authors.affiliation: Meningococcal Reference Unit, Public Health England, Manchester, United Kingdom.
authors.forename: Edward B
authors.initials: EB
authors.lastname: Kaczmarski
authors.affiliation: National Institute for Biological Standards and Control, Potters Bar, United Kingdom.
authors.forename: Ian
authors.initials: I
authors.lastname: Feavers
authors.affiliation: Department of Genetics, University of Leicester, Leicester, United Kingdom.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Bayliss
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology; D016470:Bacteremia; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D055786:Gene Knockout Techniques; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007501:Iron / Q000378:metabolism; D008585:Meningitis, Meningococcal / Q000276:immunology* / Q000382:microbiology*; D051379:Mice; D009154:Mutation; D009345:Neisseria meningitidis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D000939:Epitopes; C094109:HmbR protein, Neisseria meningitidis; C099188:HupA protein, bacteria; D007106:Immune Sera; D011956:Receptors, Cell Surface; D007501:Iron