This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow E-mail this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ASM journals
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Awad, M. M.
Right arrow Articles by Rood, J. I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Awad, M. M.
Right arrow Articles by Rood, J. I.

 Previous Article  |  Next Article 

Infection and Immunity, December 2001, p. 7904-7910, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7904-7910.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Synergistic Effects of Alpha-Toxin and Perfringolysin O in Clostridium perfringens-Mediated Gas Gangrene

Milena M. Awad,1 Darren M. Ellemor,2 Richard L. Boyd,2 John J. Emmins,2 and Julian I. Rood1,*

Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, Victoria 3800,1 and Department of Pathology and Immunology, Monash University Medical School, Alfred Hospital, Prahran 3181,2 Australia

Received 8 June 2001/Returned for modification 24 July 2001/Accepted 24 August 2001

To examine the synergistic effects of alpha-toxin and perfringolysin O in clostridial myonecrosis, homologous recombination was used to construct an alpha-toxin deficient derivative of a perfringolysin O mutant of Clostridium perfringens. The subsequent strain was complemented with separate plasmids that carried the alpha-toxin structural gene (plc), the perfringolysin O gene (pfoA), or both toxin genes, and the resultant isogenic strains were examined in a mouse myonecrosis model. Synergistic effects were clearly observed in these experiments. Infection with the control strain, which did not produce either toxin, resulted in very minimal gross pathological changes, whereas the isogenic strain that was reconstituted for both toxins produced a pathology that was clearly more severe than when alpha-toxin alone was reconstituted. These changes were most apparent in the rapid spread of the disease, the gross pathology of the footpad and in the rate at which the mice had to be euthanatized for ethical reasons. Elimination of both alpha-toxin and perfringolysin O production removed most of the histopathological features typical of clostridial myonecrosis. These effects were restored when the mutant was complemented with the alpha-toxin structural gene, but reconstituting only perfringolysin O activity produced vastly different results, with regions of coagulative necrosis, apparently enhanced by vascular disruption, being observed. Reconstitution of both alpha-toxin and perfringolysin O activity produced histopathology most similar to that observed with the alpha-toxin reconstituted strain. The spreading of myonecrosis was very rapid in these tissues, and coagulative necrosis appeared to be restricted to the lumen of the blood vessels. The results of these virulence experiments clearly support the hypothesis that alpha-toxin and perfringolysin O have a synergistic effect in the pathology of gas gangrene.

* Corresponding author. Mailing address: Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, P.O. Box 53, Victoria 3800, Australia. Phone: (613) 99054825. Fax: (613) 99054811. E-mail: Julian.Rood{at}med.monash.edu.au.

Infection and Immunity, December 2001, p. 7904-7910, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7904-7910.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Chiarezza, M., Lyras, D., Pidot, S. J., Flores-Diaz, M., Awad, M. M., Kennedy, C. L., Cordner, L. M., Phumoonna, T., Poon, R., Hughes, M. L., Emmins, J. J., Alape-Giron, A., Rood, J. I. (2009). The NanI and NanJ Sialidases of Clostridium perfringens Are Not Essential for Virulence. Infect. Immun. 77: 4421-4428 [Abstract] [Full Text]  
  • Kennedy, C. L., Lyras, D., Cordner, L. M., Melton-Witt, J., Emmins, J. J., Tweten, R. K., Rood, J. I. (2009). Pore-Forming Activity of Alpha-Toxin Is Essential for Clostridium septicum-Mediated Myonecrosis. Infect. Immun. 77: 943-951 [Abstract] [Full Text]  
  • Kulkarni, R. R., Parreira, V. R., Sharif, S., Prescott, J. F. (2007). Immunization of Broiler Chickens against Clostridium perfringens-Induced Necrotic Enteritis. CVI 14: 1070-1077 [Abstract] [Full Text]  
  • Rekha, A., Gopalan, T.R. (2007). Civilian Gas Gangrene: A Clinical Challenge. INT J LOW EXTREM WOUNDS 6: 98-101 [Abstract]  
  • Keyburn, A. L., Sheedy, S. A., Ford, M. E., Williamson, M. M., Awad, M. M., Rood, J. I., Moore, R. J. (2006). Alpha-Toxin of Clostridium perfringens Is Not an Essential Virulence Factor in Necrotic Enteritis in Chickens. Infect. Immun. 74: 6496-6500 [Abstract] [Full Text]  
  • Fisher, D. J., Fernandez-Miyakawa, M. E., Sayeed, S., Poon, R., Adams, V., Rood, J. I., Uzal, F. A., McClane, B. A. (2006). Dissecting the Contributions of Clostridium perfringens Type C Toxins to Lethality in the Mouse Intravenous Injection Model. Infect. Immun. 74: 5200-5210 [Abstract] [Full Text]  
  • Myers, G. S.A., Rasko, D. A., Cheung, J. K., Ravel, J., Seshadri, R., DeBoy, R. T., Ren, Q., Varga, J., Awad, M. M., Brinkac, L. M., Daugherty, S. C., Haft, D. H., Dodson, R. J., Madupu, R., Nelson, W. C., Rosovitz, M.J., Sullivan, S. A., Khouri, H., Dimitrov, G. I., Watkins, K. L., Mulligan, S., Benton, J., Radune, D., Fisher, D. J., Atkins, H. S., Hiscox, T., Jost, B. H., Billington, S. J., Songer, J. G., McClane, B. A., Titball, R. W., Rood, J. I., Melville, S. B., Paulsen, I. T. (2006). Skewed genomic variability in strains of the toxigenic bacterial pathogen, Clostridium perfringens. Genome Res 16: 1031-1040 [Abstract] [Full Text]  
  • Kanuck, D. M., Zgonis, T., Jolly, G. P. (2006). Necrotizing Fasciitis in a Patient with Type 2 Diabetes Mellitus. J. Am. Podiatr. Med. Assoc. 96: 67-72 [Abstract] [Full Text]  
  • Chen, Y., McClane, B. A., Fisher, D. J., Rood, J. I., Gupta, P. (2005). Construction of an Alpha Toxin Gene Knockout Mutant of Clostridium perfringens Type A by Use of a Mobile Group II Intron. Appl. Environ. Microbiol. 71: 7542-7547 [Abstract] [Full Text]  
  • Sayeed, S., Fernandez-Miyakawa, M. E., Fisher, D. J., Adams, V., Poon, R., Rood, J. I., Uzal, F. A., McClane, B. A. (2005). Epsilon-Toxin Is Required for Most Clostridium perfringens Type D Vegetative Culture Supernatants To Cause Lethality in the Mouse Intravenous Injection Model. Infect. Immun. 73: 7413-7421 [Abstract] [Full Text]  
  • O'Brien, D. K., Melville, S. B. (2004). Effects of Clostridium perfringens Alpha-Toxin (PLC) and Perfringolysin O (PFO) on Cytotoxicity to Macrophages, on Escape from the Phagosomes of Macrophages, and on Persistence of C. perfringens in Host Tissues. Infect. Immun. 72: 5204-5215 [Abstract] [Full Text]  
  • Varga, J., Stirewalt, V. L., Melville, S. B. (2004). The CcpA Protein Is Necessary for Efficient Sporulation and Enterotoxin Gene (cpe) Regulation in Clostridium perfringens. J. Bacteriol. 186: 5221-5229 [Abstract] [Full Text]  
  • Cheung, J. K., Dupuy, B., Deveson, D. S., Rood, J. I. (2004). The Spatial Organization of the VirR Boxes Is Critical for VirR-Mediated Expression of the Perfringolysin O Gene, pfoA, from Clostridium perfringens. J. Bacteriol. 186: 3321-3330 [Abstract] [Full Text]  
  • Awad, M. M., Rood, J. I. (2002). Perfringolysin O Expression in Clostridium perfringens Is Independent of the Upstream pfoR Gene. J. Bacteriol. 184: 2034-2038 [Abstract] [Full Text]  


Copyright © 2010 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»