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Infection and Immunity, August 2001, p. 4726-4733, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4726-4733.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Haemophilus ducreyi Inhibits Phagocytosis by U-937 Cells, a Human Macrophage-Like Cell Line

Gwendolyn E. Wood,dagger Susan M. Dutro, and Patricia A. Totten*

Department of Medicine, Division of Infectious Diseases, University of Washington, Seattle, Washington

Received 8 January 2001/Returned for modification 2 March 2001/Accepted 19 April 2001

Haemophilus ducreyi is a gram-negative obligate human pathogen that causes the genital ulcer disease chancroid. Chancroid lesions are deep necrotic ulcers with an immune cell infiltrate that includes macrophages. Despite the presence of these phagocytic cells, chancroid ulcers can persist for months and live H. ducreyi can be isolated from these lesions. To analyze the interaction of H. ducreyi with macrophages, we investigated the ability of H. ducreyi strain 35000 to adhere to, invade, and survive within U-937 cells, a human macrophage-like cell line. We found that although H. ducreyi strain 35000 adhered efficiently to U-937 cells, few bacteria were internalized, suggesting that H. ducreyi avoids phagocytosis by human macrophages. The few bacteria that were phagocytosed in these experiments were rapidly killed. We also found that H. ducreyi inhibits the phagocytosis of a secondary target (opsonized sheep red blood cells). Antiphagocytic activity was found in logarithmic, stationary-phase, and plate-grown cultures and was associated with whole, live bacteria but not with heat-killed cultures, sonicates, or culture supernatants. Phagocytosis was significantly inhibited after a 15-min exposure to H. ducreyi, and a multiplicity of infection of approximately 1 CFU per macrophage was sufficient to cause a significant reduction in phagocytosis by U-937 cells. Finally, all of nine H. ducreyi strains tested were antiphagocytic, suggesting that this is a common virulence mechanism for this organism. This finding suggests a mechanism by which H. ducreyi avoids killing and clearance by macrophages in chancroid lesions and inguinal lymph nodes.

* Corresponding author. Mailing address: Division of Infectious Diseases, Box 359779, Harborview Medical Center, 325 Ninth Ave., Seattle, WA 98104. Phone: (206) 341-5350. Fax: (206) 341-5363. E-mail: patotten{at}u.washington.edu.

dagger Present address: Department of Microbiology, Box 357242, University of Washington, Seattle, WA 98195.

Infection and Immunity, August 2001, p. 4726-4733, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4726-4733.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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