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Infection and Immunity, May 2000, p. 2602-2607, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Evaluation of an Isogenic Major Outer Membrane Protein-Deficient Mutant in the Human Model of Haemophilus ducreyi Infection

Robert E. Throm,1 Jaffar A. Al-Tawfiq,2 Kate R. Fortney,2 Barry P. Katz,2 Antoinette F. Hood,3,4 Clive A. Slaughter,5 Eric J. Hansen,6 and Stanley M. Spinola1,2,4,*

Departments of Microbiology and Immunology,1 Medicine,2 Dermatology,3 and Pathology and Laboratory Medicine,4 School of Medicine, Indiana University, Indianapolis, Indiana 46202, and Departments of Biochemistry5 and Microbiology,6 University of Texas Southwestern Medical Center, Dallas, Texas 75235

Received 3 November 1999/Returned for modification 21 December 1999/Accepted 1 February 2000

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.

* Corresponding author. Mailing address: Department of Medicine, 435 Emerson Hall, 545 Barnhill Dr., Indiana University, Indianapolis, IN 46202-5124. Phone: (317) 274-1427. Fax: (317) 274-1587. E-mail: sspinola{at}iupui.edu.

Infection and Immunity, May 2000, p. 2602-2607, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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