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Infection and Immunity, November 2005, p. 7495-7501, Vol. 73, No. 11
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.11.7495-7501.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Murine Macrophages Kill the Vegetative Form of Bacillus anthracis

Tae Jin Kang,1 Matthew J. Fenton,2 Matthew A. Weiner,3 Stephen Hibbs,4 Subhendu Basu,2 Les Baillie,3,4 and Alan S. Cross1*

Center for Vaccine Development,1 Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of Maryland School of Medicine, Baltimore, Maryland,2 Biological Defense Research Directorate, Naval Medical Research Center, Silver Spring, Maryland,3 Medical Biotechnology Center, University of Maryland Biotechnology Institute, Baltimore, Maryland4

Received 28 June 2005/ Returned for modification 29 July 2005/ Accepted 18 August 2005

Anti-protective antigen antibody was reported to enhance macrophage killing of ingested Bacillus anthracis spores, but it was unclear whether the antibody-mediated macrophage killing mechanism was directed against the spore itself or the vegetative form emerging from the ingested and germinating spore. To address this question, we compared the killing of germination-proficient (gp) and germination-deficient ({Delta}gerH) Sterne 34F2 strain spores by murine peritoneal macrophages. While macrophages similarly ingested both spores, only gp Sterne was killed at 5 h (0.37 log kill). Pretreatment of macrophages with gamma interferon (IFN-{gamma}) or opsonization with immunoglobulin G (IgG) isolated from a subject immunized with an anthrax vaccine enhanced the killing of Sterne to 0.49 and 0.73 log, respectively, but the combination of IFN-{gamma} and IgG was no better than either treatment alone. Under no condition was there killing of {Delta}gerH spores. To examine the ability of the exosporium to protect spores from macrophages, we compared the macrophage-mediated killing of nonsonicated (exosporium+) and sonicated (exosporium–) Sterne 34F2 spores. More sonicated spores than nonsonicated spores were killed at 5 h (0.98 versus 0.37 log kill, respectively). Pretreatment with IFN-{gamma} increased the sonicated spore killing to 1.39 log. However, the opsonization with IgG was no better than no treatment or pretreatment with IFN-{gamma}. We conclude that macrophages appear unable to kill the spore form of B. anthracis and that the exosporium may play a role in the protection of spores from macrophages.

* Corresponding author. Mailing address: Center for Vaccine Development, Department of Medicine, University of Maryland, 685 W. Baltimore Street, HSF I-480, Baltimore, MD 21201. Phone: (410) 706-6499. Fax: (410) 706-6205. E-mail: across{at}medicine.umaryland.edu.

Editor: D. L. Burns

Infection and Immunity, November 2005, p. 7495-7501, Vol. 73, No. 11
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.11.7495-7501.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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