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Infection and Immunity, May 2007, p. 2351-2358, Vol. 75, No. 5
0019-9567/07/$08.00+0     doi:10.1128/IAI.01982-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Role of Bacillus anthracis Spore Structures in Macrophage Cytokine Responses{triangledown}

Subhendu Basu,1* Tae Jin Kang,1 Wilbur H. Chen,1 Matthew J. Fenton,2,{dagger} Les Baillie,3,4 Steve Hibbs,3 and Alan S. Cross1

Center for Vaccine Development (CVD), Department of Medicine, University of Maryland School of Medicine, Baltimore, Maryland,1 Mucosal Biology Research Center, University of Maryland School of Medicine, Baltimore, Maryland,2 Medical Biotechnology Center, University of Maryland Biotechnology Institute (UMBI), Baltimore, Maryland,3 Biological Defense Research Directorate, Naval Medical Research Center (NMRC), Silver Spring, Maryland4

Received 18 December 2006/ Returned for modification 29 January 2007/ Accepted 15 February 2007

The innate immune response of macrophages (M{phi}) to spores, the environmentally acquired form of Bacillus anthracis, is poorly characterized. We therefore examined the early M{phi} cytokine response to B. anthracis spores, before germination. M{phi} were exposed to bacilli and spores of Sterne strain 34F2 and its congenic nongerminating mutant ({Delta}gerH), and cytokine expression was measured by real-time PCR and an enzyme-linked immunosorbent assay. The exosporium spore layer was retained (exo+) or removed by sonication (exo–). Spores consistently induced a strong cytokine response, with the exo– spores eliciting a two- to threefold-higher response than exo+ spores. The threshold for interleukin-1ß (IL-1ß) production by wild-type M{phi} was significantly lower than that required for tumor necrosis factor alpha expression. Cytokine production was largely dependent on MyD88, suggesting Toll-like receptor involvement; however, the expression of beta interferon in MyD88–/– M{phi} suggests involvement of a MyD88-independent pathway. We conclude that (i) the B. anthracis spore is not immunologically inert, (ii) the exosporium masks epitopes recognized by the M{phi}, (iii) the M{phi} cytokine response to B. anthracis involves multiple pattern recognition receptors and signaling pathways, and (iv) compared to other cytokines, IL-1ß is expressed at a lower spore concentration.

* 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) 706-5328. Fax: (410) 706-6205. E-mail: sbasu1{at}medicine.umaryland.edu

{triangledown} Published ahead of print on 5 March 2007.

Editor: D. L. Burns

{dagger} Present address: National Institute of Allergy and Infectious Diseases, Bethesda, MD.

Infection and Immunity, May 2007, p. 2351-2358, Vol. 75, No. 5
0019-9567/07/$08.00+0     doi:10.1128/IAI.01982-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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