title: Outer membrane porin M35 of Moraxella catarrhalis mediates susceptibility to aminopenicillins.
abstract: BACKGROUND
  The outer membrane protein M35 is a conserved porin of type 1 strains of the respiratory pathogen Moraxella catarrhalis. It was previously shown that M35 is involved in the uptake of essential nutrients required for bacterial growth and for nasal colonization in mice. The aim of this study was (i) to characterize the potential roles of M35 in the host-pathogen interactions considering the known multifunctionality of porins and (ii) to characterize the degree of conservation in the phylogenetic older subpopulation (type 2) of M. catarrhalis.
  
  
  RESULTS
  Isogenic m35 mutants of the type 1 strains O35E, 300 and 415 were tested for their antimicrobial susceptibility against 15 different agents. Differences in the MIC (Minimum Inhibitory Concentration) between wild-type and mutant strains were found for eight antibiotics. For ampicillin and amoxicillin, we observed a statistically significant 2.5 to 2.9-fold MIC increase (p < 0.03) in the m35 mutants. Immunoblot analysis demonstrated that human saliva contains anti-M35 IgA. Wild-type strains and their respective m35 mutants were indistinguishable with respect to the phenotypes of autoagglutination, serum resistance, iron acquisition from human lactoferrin, adherence to and invasion of respiratory tract epithelial cells, and proinflammatory stimulation of human monocytes. DNA sequencing of m35 from the phylogenetic subpopulation type 2 strain 287 revealed 94.2% and 92.8% identity on the DNA and amino acid levels, respectively, in comparison with type 1 strains.
  
  
  CONCLUSIONS
  The increase in MIC for ampicillin and amoxicillin, respectively, in the M35-deficient mutants indicates that this porin affects the outer membrane permeability for aminopenicillins in a clinically relevant manner. The presence of IgA antibodies in healthy human donors indicates that M35 is expressed in vivo and recognized as a mucosal antigen by the human host. However, immunoblot analysis of human saliva suggests the possibility of antigenic variation of immunoreactive epitopes, which warrants further analysis before M35 can be considered a potential vaccine candidate.
authors.affiliation: Institute for Infectious Diseases, University of Bern, CH-3010 Bern, Switzerland. marion.jetter@ifik.unibe.ch
authors.forename: Marion
authors.initials: M
authors.lastname: Jetter
authors.forename: Nadja
authors.initials: N
authors.lastname: Heiniger
authors.forename: Violeta
authors.initials: V
authors.lastname: Spaniol
authors.forename: Rolf
authors.initials: R
authors.lastname: Troller
authors.forename: André
authors.initials: A
authors.lastname: Schaller
authors.forename: Christoph
authors.initials: C
authors.lastname: Aebi
mesh_terms: D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D054884:Host-Pathogen Interactions; D006801:Humans; D007070:Immunoglobulin A / Q000276:immunology; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D001936:Moraxella catarrhalis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D016254:Mutagenesis, Insertional; D010406:Penicillins / Q000494:pharmacology*; D010802:Phylogeny; D018272:Porins / Q000235:genetics / Q000378:metabolism*; D012463:Saliva / Q000276:immunology; D014774:Virulence
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D007070:Immunoglobulin A; D010406:Penicillins; D018272:Porins