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Infection and Immunity, December 1999, p. 6335-6340, Vol. 67, No. 12
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Expression of Sialylated or Paragloboside-Like Lipooligosaccharides Are Not Required for Pustule Formation by Haemophilus ducreyi in Human Volunteers

Royden S. Young,1 Kate Fortney,1 Jennifer C. Haley,2 Antoinette F. Hood,1,2 Anthony A. Campagnari,3 Jing Wang,4 Joel A. Bozue,4 Robert S. Munson Jr.,4,5 and Stanley M. Spinola1,6,7,*

Departments of Medicine,1 Microbiology and Immunology,6 Pathology and Laboratory Medicine,7 and Dermatology,2 Indiana University School of Medicine, Indianapolis, Indiana 46202; Department of Microbiology, State University of New York at Buffalo, New York 142143; and Children's Research Institute4 and Department of Pediatrics and Department of Medical Microbiology and Immunology,5 Ohio State University, Columbus, Ohio 43205-2696

Received 1 July 1999/Returned for modification 16 August 1999/Accepted 13 September 1999

The lipooligosaccharide (LOS) of Haemophilus ducreyi, the etiologic agent of chancroid, chemically and immunologically resembles human glycosphingolipid antigens. To test whether LOS that contains paragloboside-like structures was required for pustule formation, an isogenic mutant (35000HP-RSM2) was constructed in losB, which encodes D-glycero-D-manno-heptosyltransferase. 35000HP-RSM2 produces a truncated LOS whose major glycoform terminates in a single glucose attached to a heptose trisaccharide core and 2-keto-3-deoxyoctulosonic acid. Five human subjects were inoculated with 35000HP and 35000HP-RSM2 in a dose-response trial. For estimated delivered doses (EDDs) of >= 25 CFU, the pustule formation rates were 80% for 35000HP and 58% for 35000HP-RSM2. Preliminary data indicated that a previously described Tn916 losB mutant made a minor glycoform that does not require DD-heptose to form the terminal N-acetyllactosamine. If 35000HP-RSM2 made this glycoform, then 35000HP-RSM2 could theoretically make a sialylated glycoform. To test whether sialylated LOS was required for pustule formation, a second trial comparing an isogenic sialyltransferase mutant (35000HP-RSM203) to 35000HP was performed in five additional subjects. For EDDs of >= 25 CFU, the pustule formation rates were 30% for both 35000HP and 35000HP-RSM203. The histopathology and recovery rates of H. ducreyi from surface cultures and biopsies obtained from mutant and parent sites in both trials were similar. These results indicate that neither the expression of a major glycoform resembling paragloboside nor sialylated LOS is required for pustule formation by H. ducreyi in humans.

* 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, December 1999, p. 6335-6340, Vol. 67, No. 12
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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