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Infection and Immunity, April 2000, p. 2309-2314, Vol. 68, No. 4
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Localization of Haemophilus ducreyi at the Pustular Stage of Disease in the Human Model of Infection

Margaret E. Bauer1,* and Stanley M. Spinola1,2,3

Departments of Medicine,1 Microbiology and Immunology,2 and Pathology and Laboratory Medicine,3 Indiana University School of Medicine, Indianapolis, Indiana 46202

Received 29 October 1999/Returned for modification 15 December 1999/Accepted 29 December 1999

To localize Haemophilus ducreyi in vivo, human subjects were experimentally infected with H. ducreyi until they developed a painful pustule or for 14 days. Lesions were biopsied, and biopsy samples were fixed in 4% paraformaldehyde, and cryosectioned. Sections were stained with polyclonal anti-H. ducreyi antiserum or H. ducreyi-specific monoclonal antibodies (MAbs) and fluorescently tagged secondary antibodies and examined by confocal microscopy. We identified H. ducreyi in 16 of 18 pustules but did not detect bacteria in the one papule examined. H. ducreyi was observed as individual cells and in clumps or chains. Staining with MAbs 2D8, 5C9, 3B9, 2C7, and 9D12 demonstrated that H. ducreyi expresses the major pilus subunit, FtpA, the 28-kDa outer membrane protein Hlp, the 18-kDa outer membrane protein PAL, and the major outer membrane protein (MOMP) or OmpA2 in vivo. By dual staining with polyclonal anti-H. ducreyi antiserum and MAbs that recognize human skin components, we observed bacteria within the neutrophilic infiltrates of all positively staining pustules and in the dermis of 10 of 16 pustules. We were unable to detect bacteria associated with keratinocytes in the samples examined. The data suggest that H. ducreyi is found primarily in association with neutrophils and in the dermis at the pustular stage of disease in the human model of infection.

* Corresponding author. Mailing address: Indiana University School of Medicine, Emerson Hall 435, 545 Barnhill Dr., Indianapolis, IN 46202. Phone: (317) 274-8143. Fax: (317) 274-1587. E-mail: mebauer{at}iupui.edu.

Infection and Immunity, April 2000, p. 2309-2314, Vol. 68, No. 4
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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