title: Evaluation of an isogenic major outer membrane protein-deficient mutant in the human model of Haemophilus ducreyi infection.
abstract: Haemophilus ducreyi expresses 2 OmpA homologs, designated MOMP and OmpA2, whose genes are arranged in tandem on the chromosome. Northern blot analysis indicated that momp and ompA2 are transcribed independently. Sequences of the momp open reading frame (ORF) lacking the transcriptional start site were amplified by PCR, and an Omega-Km2 cassette was ligated into the ORF. A plasmid containing this construction was electroporated into H. ducreyi 35000HP, and an isogenic MOMP-deficient mutant (35000HP-SMS2) was generated by allele exchange. In Southern blotting, 35000HP-SMS2 contained one copy of the Omega-Km2 cassette in momp. 35000HP and 35000HP-SMS2 had similar outer membrane protein (OMP) and lipooligosaccharide profiles and growth rates except for up-regulation of a putative porin protein in the mutant. Five subjects were inoculated with three doses of live 35000HP-SMS2 on one arm and two doses of live 35000HP and one dose of a heat-killed control on the other arm in a double-blind escalating dose-response trial. Pustules developed at 7 of 10 sites inoculated with 35000HP and at 6 of 15 sites inoculated with 35000HP-SMS2 (P = 0.14). 35000HP and 35000HP-SMS2 were recovered at similar rates from daily surface cultures and semiquantitative cultures. The data suggest that expression of MOMP is not required for pustule formation by H. ducreyi in the human model of infection.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, Indiana University, Indianapolis, Indiana 46202, USA.
authors.forename: R E
authors.initials: RE
authors.lastname: Throm
authors.forename: J A
authors.initials: JA
authors.lastname: Al-Tawfiq
authors.forename: K R
authors.initials: KR
authors.lastname: Fortney
authors.forename: B P
authors.initials: BP
authors.lastname: Katz
authors.forename: A F
authors.initials: AF
authors.lastname: Hood
authors.forename: C A
authors.initials: CA
authors.lastname: Slaughter
authors.forename: E J
authors.initials: EJ
authors.lastname: Hansen
authors.forename: S M
authors.initials: SM
authors.lastname: Spinola
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000502:physiology*; D002602:Chancroid / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D005260:Female; D017353:Gene Deletion; D006191:Haemophilus ducreyi / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D006801:Humans; D008954:Models, Biological; D008969:Molecular Sequence Data; D010641:Phenotype; D014158:Transcription, Genetic
chemical_list: D001425:Bacterial Outer Membrane Proteins; C104640:OmpA2 protein, Haemophilus decreyi; C078911:major outer membrane protein, Haemophilus