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Infection and Immunity, February 2009, p. 749-755, Vol. 77, No. 2
0019-9567/09/$08.00+0     doi:10.1128/IAI.00764-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Association of Bacillus anthracis Capsule with Lethal Toxin during Experimental Infection{triangledown}

J. W. Ezzell,1 T. G. Abshire,1 R. Panchal,3 D. Chabot,2 S. Bavari,2 E. K. Leffel,4 B. Purcell,2 A. M. Friedlander,5 and W. J. Ribot2*

Diagnostic Systems Division,1 Bacteriology Division,2 Toxinology Division,3 National Biodefense Analysis and Countermeasures Center,4 Headquarters, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 217025

Received 18 June 2008/ Returned for modification 26 July 2008/ Accepted 27 November 2008

Bacillus anthracis lethal toxin (LT) was characterized in plasma from infected African Green monkeys, rabbits, and guinea pigs. In all cases, during the terminal phase of infection only the protease-activated 63-kDa form of protective antigen (PA63) and the residual 20-kDa fragment (PA20) were detected in the plasma. No uncut PA with a molecular mass of 83 kDa was detected in plasma from toxemic animals during the terminal stage of infection. PA63 was largely associated with lethal factor (LF), forming LT. Characterization of LT by Western blotting, capture enzyme-linked immunosorbent assay, and size exclusion chromatography revealed that the antiphagocytic poly-{gamma}-D-glutamic acid ({gamma}-DPGA) capsule released from B. anthracis bacilli was associated with LT in animal blood in variable amounts. While the nature of this in vivo association is not understood, we were able to determine that a portion of these LT/{gamma}-DPGA complexes retained LF protease activity. Our findings suggest that the in vivo LT complexes differ from in vitro-produced LT and that including {gamma}-DPGA when examining the effects of LT on specific immune cells in vitro may reveal novel and important roles for {gamma}-DPGA in anthrax pathogenesis.

* Corresponding author. Mailing address: Department of Bacteriology, United States Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702. Phone: (301) 619-4886. Fax: (301) 619-2153. E-mail: Wilson.ribot{at}amedd.army.mil

{triangledown} Published ahead of print on 8 December 2008.

Editor: S. R. Blanke

Infection and Immunity, February 2009, p. 749-755, Vol. 77, No. 2
0019-9567/09/$08.00+0     doi:10.1128/IAI.00764-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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