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
Services
Right arrow Similar articles in this journal
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 Stevens, M. K.
Right arrow Articles by Hansen, E. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Stevens, M. K.
Right arrow Articles by Hansen, E. J.

 Previous Article  |  Next Article 

Infection and Immunity, August 1999, p. 3900-3908, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Characterization of a Haemophilus ducreyi Mutant Deficient in Expression of Cytolethal Distending Toxin

Marla K. Stevens,1 Jo L. Latimer,1 Sheryl R. Lumbley,1 Christine K. Ward,1 Leslie D. Cope,1 Teresa Lagergard,2 and Eric J. Hansen1,*

Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas 75235-9048,1 and Department of Medical Microbiology and Immunology, University of Goteborg, S-413 46 Goteborg, Sweden2

Received 5 February 1999/Returned for modification 18 March 1999/Accepted 6 May 1999

Haemophilus ducreyi expresses a soluble cytolethal distending toxin (CDT) that kills HeLa, HEp-2, and other human epithelial cells in vitro. H. ducreyi CDT activity is encoded by a three-gene cluster (cdtABC), and antibody to the cdtC gene product can neutralize CDT activity in vitro (L. D. Cope, S. R. Lumbley, J. L. Latimer, J. Klesney-Tait, M. K. Stevens, L. S. Johnson, M. Purven, R. S. Munson, Jr., T. Lagergard, J. D. Radolf, and E. J. Hansen, Proc. Natl. Acad. Sci. USA 94:4056-4061, 1997). Culture supernatant fluid from a recombinant Escherichia coli strain containing the H. ducreyi cdtABC gene cluster readily killed both HeLa cells and HaCaT keratinocytes and had a modest inhibitory effect on the growth of human foreskin fibroblasts. Insertional inactivation of the cdtC gene in this recombinant E. coli strain eliminated the ability of this strain to kill HeLa cells and HaCaT keratinocytes. This mutated H. ducreyi cdtABC gene cluster was used to construct an isogenic H. ducreyi cdtC mutant. Monoclonal antibodies against the H. ducreyi CdtA, CdtB, and CdtC proteins were used to characterize protein expression by this cdtC mutant. Culture supernatant fluid from this H. ducreyi cdtC mutant did not detectably affect any of the human cells used in this study. The presence of the wild-type H. ducreyi cdtC gene in trans in this H. ducreyi mutant restored its ability to express a CDT that killed both HeLa cells and HaCaT keratinocytes. The isogenic H. ducreyi cdtC mutant was shown to be as virulent as its wild-type parent strain in the temperature-dependent rabbit model for experimental chancroid. Lack of expression of the H. ducreyi CdtC protein also did not affect the ability of this H. ducreyi mutant to survive in the skin of rabbits.

* Corresponding author. Mailing address: Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235-9048. Phone: (214) 648-5974. Fax: (214) 648-5905. E-mail: hansen01{at}utsw.swmed.edu.

Infection and Immunity, August 1999, p. 3900-3908, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Bauer, M. E., Fortney, K. R., Harrison, A., Janowicz, D. M., Munson, R. S., Spinola, S. M. (2008). Identification of Haemophilus ducreyi genes expressed during human infection. Microbiology 154: 1152-1160 [Abstract] [Full Text]  
  • Banks, K. E., Humphreys, T. L., Li, W., Katz, B. P., Wilkes, D. S., Spinola, S. M. (2007). Haemophilus ducreyi Partially Activates Human Myeloid Dendritic Cells. Infect. Immun. 75: 5678-5685 [Abstract] [Full Text]  
  • Cao, L., Volgina, A., Korostoff, J., DiRienzo, J. M. (2006). Role of Intrachain Disulfides in the Activities of the CdtA and CdtC Subunits of the Cytolethal Distending Toxin of Actinobacillus actinomycetemcomitans. Infect. Immun. 74: 4990-5002 [Abstract] [Full Text]  
  • Ge, Z., Feng, Y., Whary, M. T., Nambiar, P. R., Xu, S., Ng, V., Taylor, N. S., Fox, J. G. (2005). Cytolethal Distending Toxin Is Essential for Helicobacter hepaticus Colonization in Outbred Swiss Webster Mice. Infect. Immun. 73: 3559-3567 [Abstract] [Full Text]  
  • Heywood, W., Henderson, B., Nair, S. P (2005). Cytolethal distending toxin: creating a gap in the cell cycle. J Med Microbiol 54: 207-216 [Abstract] [Full Text]  
  • Mohd-Zain, Z., Turner, S. L., Cerdeno-Tarraga, A. M., Lilley, A. K., Inzana, T. J., Duncan, A. J., Harding, R. M., Hood, D. W., Peto, T. E., Crook, D. W. (2004). Transferable Antibiotic Resistance Elements in Haemophilus influenzae Share a Common Evolutionary Origin with a Diverse Family of Syntenic Genomic Islands. J. Bacteriol. 186: 8114-8122 [Abstract] [Full Text]  
  • Young, V. B., Knox, K. A., Pratt, J. S., Cortez, J. S., Mansfield, L. S., Rogers, A. B., Fox, J. G., Schauer, D. B. (2004). In Vitro and In Vivo Characterization of Helicobacter hepaticus Cytolethal Distending Toxin Mutants. Infect. Immun. 72: 2521-2527 [Abstract] [Full Text]  
  • Shenker, B. J., Besack, D., McKay, T., Pankoski, L., Zekavat, A., Demuth, D. R. (2004). Actinobacillus actinomycetemcomitans Cytolethal Distending Toxin (Cdt): Evidence That the Holotoxin Is Composed of Three Subunits: CdtA, CdtB, and CdtC. J. Immunol. 172: 410-417 [Abstract] [Full Text]  
  • Spinola, S. M., Fortney, K. R., Katz, B. P., Latimer, J. L., Mock, J. R., Vakevainen, M., Hansen, E. J. (2003). Haemophilus ducreyi Requires an Intact flp Gene Cluster for Virulence in Humans. Infect. Immun. 71: 7178-7182 [Abstract] [Full Text]  
  • Deng, K., Hansen, E. J. (2003). A CdtA-CdtC Complex Can Block Killing of HeLa Cells by Haemophilus ducreyi Cytolethal Distending Toxin. Infect. Immun. 71: 6633-6640 [Abstract] [Full Text]  
  • Kulkarni, K, Lewis, D A, Ison, C A (2003). Expression of the cytolethal distending toxin in a geographically diverse collection of Haemophilus ducreyi clinical isolates. Sex. Transm. Infect. 79: 294-297 [Abstract] [Full Text]  
  • Svensson, L. A., Henning, P., Lagergard, T. (2002). The Cytolethal Distending Toxin of Haemophilus ducreyi Inhibits Endothelial Cell Proliferation. Infect. Immun. 70: 2665-2669 [Abstract] [Full Text]  
  • Spinola, S. M., Bauer, M. E., Munson, R. S. Jr. (2002). Immunopathogenesis of Haemophilus ducreyi Infection (Chancroid). Infect. Immun. 70: 1667-1676 [Full Text]  
  • Henderson, B. (2002). Oral bacterial disease and the science of cellular conversation. JRSM 95: 77-80 [Full Text]  
  • AHMED, H.J., SVENSSON, L.A., COPE, L.D., LATIMER, J.L., HANSEN, E.J., AHLMAN, K., BAYAT-TURK, J., KLAMER, D., LAGERGARD, T. (2001). Prevalence of cdtABC genes encoding cytolethal distending toxin among Haemophilus ducreyi and Actinobacillus actinomycetemcomitans strains. J Med Microbiol 50: 860-864 [Abstract] [Full Text]  
  • Lewis, D. A., Stevens, M. K., Latimer, J. L., Ward, C. K., Deng, K., Blick, R., Lumbley, S. R., Ison, C. A., Hansen, E. J. (2001). Characterization of Haemophilus ducreyi cdtA, cdtB, and cdtC Mutants in In Vitro and In Vivo Systems. Infect. Immun. 69: 5626-5634 [Abstract] [Full Text]  
  • Akifusa, S., Poole, S., Lewthwaite, J., Henderson, B., Nair, S. P. (2001). Recombinant Actinobacillus actinomycetemcomitans Cytolethal Distending Toxin Proteins Are Required To Interact To Inhibit Human Cell Cycle Progression and To Stimulate Human Leukocyte Cytokine Synthesis. Infect. Immun. 69: 5925-5930 [Abstract] [Full Text]  
  • Wood, G. E., Dutro, S. M., Totten, P. A. (2001). Haemophilus ducreyi Inhibits Phagocytosis by U-937 Cells, a Human Macrophage-Like Cell Line. Infect. Immun. 69: 4726-4733 [Abstract] [Full Text]  
  • Shenker, B. J., Hoffmaster, R. H., Zekavat, A., Yamaguchi, N., Lally, E. T., Demuth, D. R. (2001). Induction of Apoptosis in Human T Cells by Actinobacillus actinomycetemcomitans Cytolethal Distending Toxin Is a Consequence of G2 Arrest of the Cell Cycle. J. Immunol. 167: 435-441 [Abstract] [Full Text]  
  • Young, R. S., Fortney, K. R., Gelfanova, V., Phillips, C. L., Katz, B. P., Hood, A. F., Latimer, J. L., Munson, R. S. Jr., Hansen, E. J., Spinola, S. M. (2001). Expression of Cytolethal Distending Toxin and Hemolysin Is Not Required for Pustule Formation by Haemophilus ducreyi in Human Volunteers. Infect. Immun. 69: 1938-1942 [Abstract] [Full Text]  
  • Shenker, B. J., Hoffmaster, R. H., McKay, T. L., Demuth, D. R. (2000). Expression of the Cytolethal Distending Toxin (Cdt) Operon in Actinobacillus actinomycetemcomitans: Evidence That the CdtB Protein Is Responsible for G2 Arrest of the Cell Cycle in Human T Cells. J. Immunol. 165: 2612-2618 [Abstract] [Full Text]  
  • PURDY, D., BUSWELL, C.M., HODGSON, A.E., McALPINE, K., HENDERSON, I., LEACH, S.A. (2000). Characterisation of cytolethal distending toxin (CDT) mutants of Campylobacter jejuni. J Med Microbiol 49: 473-479 [Abstract] [Full Text]  
  • Young, V. B., Knox, K. A., Schauer, D. B. (2000). Cytolethal Distending Toxin Sequence and Activity in the Enterohepatic Pathogen Helicobacter hepaticus. Infect. Immun. 68: 184-191 [Abstract] [Full Text]  
  • Gelfanova, V., Hansen, E. J., Spinola, S. M. (1999). Cytolethal Distending Toxin of Haemophilus ducreyi Induces Apoptotic Death of Jurkat T Cells. Infect. Immun. 67: 6394-6402 [Abstract] [Full Text]  
  • Cortes-Bratti, X., Karlsson, C., Lagergard, T., Thelestam, M., Frisan, T. (2001). The Haemophilus ducreyi Cytolethal Distending Toxin Induces Cell Cycle Arrest and Apoptosis via the DNA Damage Checkpoint Pathways. J. Biol. Chem. 276: 5296-5302 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»