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 Allewelt, M.
Right arrow Articles by Pier, G. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Allewelt, M.
Right arrow Articles by Pier, G. B.

 Previous Article  |  Next Article 

Infection and Immunity, July 2000, p. 3998-4004, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Acquisition of Expression of the Pseudomonas aeruginosa ExoU Cytotoxin Leads to Increased Bacterial Virulence in a Murine Model of Acute Pneumonia and Systemic Spread

Markus Allewelt,dagger Fadie T. Coleman, Martha Grout, Gregory P. Priebe, and Gerald B. Pier*

Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115

Received 7 February 2000/Returned for modification 8 March 2000/Accepted 24 March 2000

Pseudomonas aeruginosa is the nosocomial bacterial pathogen most commonly isolated from the respiratory tract. Animal models of this infection are extremely valuable for studies of virulence and immunity. We thus evaluated the utility of a simple model of acute pneumonia for analyzing P. aeruginosa virulence by characterizing the course of bacterial infection in BALB/c mice following application of bacteria to the nares of anesthetized animals. Bacterial aspiration into the lungs was rapid, and 67 to 100% of the inoculum could be recovered within minutes from the lungs, with 0.1 to 1% of the inoculum found intracellularly shortly after infection. At later time points up to 10% of the bacteria were intracellular, as revealed by gentamicin exclusion assays on single-cell suspensions of infected lungs. Expression of exoenzyme U (ExoU) by P. aeruginosa is associated with a cytotoxic effect on epithelial cells in vitro and virulence in animal models. Insertional mutations in the exoU gene confer a noncytotoxic phenotype on mutant strains and decrease virulence for animals. We used the model of acute pneumonia to determine whether introduction of the exoU gene into noncytotoxic strains of P. aeruginosa lacking this gene affected virulence. Seven phenotypically noncytotoxic P. aeruginosa strains were transformed with pUCP19exoUspcU which carries the exoU gene and its associated chaperone. Three of these strains became cytotoxic to cultured epithelial cells in vitro. These strains all secreted ExoU, as confirmed by detection of the ExoU protein with specific antisera. The 50% lethal dose of exoU-expressing strains was significantly lower for all three P. aeruginosa isolates carrying plasmid pUCP19exoUspcU than for the isogenic exoU-negative strains. mRNA specific for ExoU was readily detected in the lungs of animals infected with the transformed P. aeruginosa strains. Introduction of the exoU gene confers a cytotoxic phenotype on some, but not all, otherwise-noncytotoxic P. aeruginosa strains and, for recombinant strains that could express ExoU, there was markedly increased virulence in a murine model of acute pneumonia and systemic spread.

* Corresponding author. Mailing address: Channing Laboratory, 181 Longwood Ave., Boston, MA 02115. Phone: (617) 525-2269. Fax: (617) 731-1541. E-mail: gpier{at}channing.harvard.edu.

dagger Present address: Krankenhaus Zehlendorf, Lungenklinik Heckeshorn, Zum Heckeshorn 33, D-14109, Berlin, Germany.

Infection and Immunity, July 2000, p. 3998-4004, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Witowski, S. E., Walker, K. A., Miller, V. L. (2008). YspM, a Newly Identified Ysa Type III Secreted Protein of Yersinia enterocolitica. J. Bacteriol. 190: 7315-7325 [Abstract] [Full Text]  
  • Zaidi, T., Pier, G. B. (2008). Prophylactic and Therapeutic Efficacy of a Fully Human Immunoglobulin G1 Monoclonal Antibody to Pseudomonas aeruginosa Alginate in Murine Keratitis Infection. Infect. Immun. 76: 4720-4725 [Abstract] [Full Text]  
  • Diaz, M. H., Shaver, C. M., King, J. D., Musunuri, S., Kazzaz, J. A., Hauser, A. R. (2008). Pseudomonas aeruginosa Induces Localized Immunosuppression during Pneumonia. Infect. Immun. 76: 4414-4421 [Abstract] [Full Text]  
  • Priebe, G. P., Walsh, R. L., Cederroth, T. A., Kamei, A., Coutinho-Sledge, Y. S., Goldberg, J. B., Pier, G. B. (2008). IL-17 Is a Critical Component of Vaccine-Induced Protection against Lung Infection by Lipopolysaccharide-Heterologous Strains of Pseudomonas aeruginosa. J. Immunol. 181: 4965-4975 [Abstract] [Full Text]  
  • El Solh, A. A., Akinnusi, M. E., Wiener-Kronish, J. P., Lynch, S. V., Pineda, L. A., Szarpa, K. (2008). Persistent Infection with Pseudomonas aeruginosa in Ventilator-associated Pneumonia. Am. J. Respir. Crit. Care Med. 178: 513-519 [Abstract] [Full Text]  
  • Ward, P. P., Mendoza-Meneses, M., Park, P. W., Conneely, O. M. (2008). Stimulus-Dependent Impairment of the Neutrophil Oxidative Burst Response in Lactoferrin-Deficient Mice. Am. J. Pathol. 172: 1019-1029 [Abstract] [Full Text]  
  • Zaidi, T., Bajmoczi, M., Zaidi, T., Golan, D. E., Pier, G. B. (2008). Disruption of CFTR-Dependent Lipid Rafts Reduces Bacterial Levels and Corneal Disease in a Murine Model of Pseudomonas aeruginosa Keratitis. IOVS 49: 1000-1009 [Abstract] [Full Text]  
  • Scarff, J. M., Goldberg, J. B. (2008). Vaccination against Pseudomonas aeruginosa Pneumonia in Immunocompromised Mice. CVI 15: 367-375 [Abstract] [Full Text]  
  • Reiniger, N., Lee, M. M., Coleman, F. T., Ray, C., Golan, D. E., Pier, G. B. (2007). Resistance to Pseudomonas aeruginosa Chronic Lung Infection Requires Cystic Fibrosis Transmembrane Conductance Regulator-Modulated Interleukin-1 (IL-1) Release and Signaling through the IL-1 Receptor. Infect. Immun. 75: 1598-1608 [Abstract] [Full Text]  
  • DiGiandomenico, A., Rao, J., Harcher, K., Zaidi, T. S., Gardner, J., Neely, A. N., Pier, G. B., Goldberg, J. B. (2007). Intranasal immunization with heterologously expressed polysaccharide protects against multiple Pseudomonas aeruginosa infections. Proc. Natl. Acad. Sci. USA 104: 4624-4629 [Abstract] [Full Text]  
  • Lee, V. T., Pukatzki, S., Sato, H., Kikawada, E., Kazimirova, A. A., Huang, J., Li, X., Arm, J. P., Frank, D. W., Lory, S. (2007). Pseudolipasin A Is a Specific Inhibitor for Phospholipase A2 Activity of Pseudomonas aeruginosa Cytotoxin ExoU. Infect. Immun. 75: 1089-1098 [Abstract] [Full Text]  
  • Wilson, P. A., Gardner, S. D., Lambie, N. M., Commans, S. A., Crowther, D. J. (2006). Characterization of the human patatin-like phospholipase family. J. Lipid Res. 47: 1940-1949 [Abstract] [Full Text]  
  • Laskowski, M. A., Kazmierczak, B. I. (2006). Mutational Analysis of RetS, an Unusual Sensor Kinase-Response Regulator Hybrid Required for Pseudomonas aeruginosa Virulence.. Infect. Immun. 74: 4462-4473 [Abstract] [Full Text]  
  • Zolfaghar, I., Evans, D. J., Ronaghi, R., Fleiszig, S. M. J. (2006). Type III Secretion-Dependent Modulation of Innate Immunity as One of Multiple Factors Regulated by Pseudomonas aeruginosa RetS. Infect. Immun. 74: 3880-3889 [Abstract] [Full Text]  
  • Rabin, S. D. P., Veesenmeyer, J. L., Bieging, K. T., Hauser, A. R. (2006). A C-Terminal Domain Targets the Pseudomonas aeruginosa Cytotoxin ExoU to the Plasma Membrane of Host Cells.. Infect. Immun. 74: 2552-2561 [Abstract] [Full Text]  
  • Zaidi, T. S., Priebe, G. P., Pier, G. B. (2006). A Live-Attenuated Pseudomonas aeruginosa Vaccine Elicits Outer Membrane Protein-Specific Active and Passive Protection against Corneal Infection. Infect. Immun. 74: 975-983 [Abstract] [Full Text]  
  • Smedley, J. G. III, Jewell, E., Roguskie, J., Horzempa, J., Syboldt, A., Stolz, D. B., Castric, P. (2005). Influence of Pilin Glycosylation on Pseudomonas aeruginosa 1244 Pilus Function. Infect. Immun. 73: 7922-7931 [Abstract] [Full Text]  
  • Ader, F., Le Berre, R., Faure, K., Gosset, P., Epaulard, O., Toussaint, B., Polack, B., Nowak, E., Viget, N. B., Kipnis, E., Guery, B. P. (2005). Alveolar Response to Pseudomonas aeruginosa: Role of the Type III Secretion System. Infect. Immun. 73: 4263-4271 [Abstract] [Full Text]  
  • Kierbel, A., Gassama-Diagne, A., Mostov, K., Engel, J. N. (2005). The Phosphoinositol-3-Kinase-Protein Kinase B/Akt Pathway Is Critical for Pseudomonas aeruginosa Strain PAK Internalization. Mol. Biol. Cell 16: 2577-2585 [Abstract] [Full Text]  
  • Koh, A. Y., Priebe, G. P., Pier, G. B. (2005). Virulence of Pseudomonas aeruginosa in a Murine Model of Gastrointestinal Colonization and Dissemination in Neutropenia. Infect. Immun. 73: 2262-2272 [Abstract] [Full Text]  
  • Bhanot, P., Schauer, K., Coppens, I., Nussenzweig, V. (2005). A Surface Phospholipase Is Involved in the Migration of Plasmodium Sporozoites through Cells. J. Biol. Chem. 280: 6752-6760 [Abstract] [Full Text]  
  • Sato, H., Feix, J. B., Hillard, C. J., Frank, D. W. (2005). Characterization of Phospholipase Activity of the Pseudomonas aeruginosa Type III Cytotoxin, ExoU. J. Bacteriol. 187: 1192-1195 [Abstract] [Full Text]  
  • Rabin, S. D. P., Hauser, A. R. (2005). Functional Regions of the Pseudomonas aeruginosa Cytotoxin ExoU. Infect. Immun. 73: 573-582 [Abstract] [Full Text]  
  • Shaver, C. M., Hauser, A. R. (2004). Relative Contributions of Pseudomonas aeruginosa ExoU, ExoS, and ExoT to Virulence in the Lung. Infect. Immun. 72: 6969-6977 [Abstract] [Full Text]  
  • Finnan, S., Morrissey, J. P., O'Gara, F., Boyd, E. F. (2004). Genome Diversity of Pseudomonas aeruginosa Isolates from Cystic Fibrosis Patients and the Hospital Environment. J. Clin. Microbiol. 42: 5783-5792 [Abstract] [Full Text]  
  • Villena, J. A., Roy, S., Sarkadi-Nagy, E., Kim, K.-H., Sul, H. S. (2004). Desnutrin, an Adipocyte Gene Encoding a Novel Patatin Domain-containing Protein, Is Induced by Fasting and Glucocorticoids: ECTOPIC EXPRESSION OF DESNUTRIN INCREASES TRIGLYCERIDE HYDROLYSIS. J. Biol. Chem. 279: 47066-47075 [Abstract] [Full Text]  
  • Pier, G. B., Boyer, D., Preston, M., Coleman, F. T., Llosa, N., Mueschenborn-Koglin, S., Theilacker, C., Goldenberg, H., Uchin, J., Priebe, G. P., Grout, M., Posner, M., Cavacini, L. (2004). Human Monoclonal Antibodies to Pseudomonas aeruginosa Alginate That Protect against Infection by Both Mucoid and Nonmucoid Strains. J. Immunol. 173: 5671-5678 [Abstract] [Full Text]  
  • Priebe, G. P., Dean, C. R., Zaidi, T., Meluleni, G. J., Coleman, F. T., Coutinho, Y. S., Noto, M. J., Urban, T. A., Pier, G. B., Goldberg, J. B. (2004). The galU Gene of Pseudomonas aeruginosa Is Required for Corneal Infection and Efficient Systemic Spread following Pneumonia but Not for Infection Confined to the Lung. Infect. Immun. 72: 4224-4232 [Abstract] [Full Text]  
  • Smith, R. S., Wolfgang, M. C., Lory, S. (2004). An Adenylate Cyclase-Controlled Signaling Network Regulates Pseudomonas aeruginosa Virulence in a Mouse Model of Acute Pneumonia. Infect. Immun. 72: 1677-1684 [Abstract] [Full Text]  
  • Banerji, S., Flieger, A. (2004). Patatin-like proteins: a new family of lipolytic enzymes present in bacteria?. Microbiology 150: 522-525 [Full Text]  
  • Phillips, R. M., Six, D. A., Dennis, E. A., Ghosh, P. (2003). In Vivo Phospholipase Activity of the Pseudomonas aeruginosa Cytotoxin ExoU and Protection of Mammalian Cells with Phospholipase A2 Inhibitors. J. Biol. Chem. 278: 41326-41332 [Abstract] [Full Text]  
  • Lee, E. J., Cowell, B. A., Evans, D. J., Fleiszig, S. M. J. (2003). Contribution of ExsA-Regulated Factors to Corneal Infection by Cytotoxic and Invasive Pseudomonas aeruginosa in a Murine Scarification Model. IOVS 44: 3892-3898 [Abstract] [Full Text]  
  • Ajayi, T., Allmond, L. R., Sawa, T., Wiener-Kronish, J. P. (2003). Single-Nucleotide-Polymorphism Mapping of the Pseudomonas aeruginosa Type III Secretion Toxins for Development of a Diagnostic Multiplex PCR System. J. Clin. Microbiol. 41: 3526-3531 [Abstract] [Full Text]  
  • Cowell, B. A., Twining, S. S., Hobden, J. A., Kwong, M. S. F., Fleiszig, S. M. J. (2003). Mutation of lasA and lasB reduces Pseudomonas aeruginosa invasion of epithelial cells. Microbiology 149: 2291-2299 [Abstract] [Full Text]  
  • Cannon, C. L., Kowalski, M. P., Stopak, K. S., Pier, G. B. (2003). Pseudomonas aeruginosa-Induced Apoptosis Is Defective in Respiratory Epithelial Cells Expressing Mutant Cystic Fibrosis Transmembrane Conductance Regulator. Am. J. Respir. Cell Mol. Bio. 29: 188-197 [Abstract] [Full Text]  
  • Wolfgang, M. C., Kulasekara, B. R., Liang, X., Boyd, D., Wu, K., Yang, Q., Miyada, C. G., Lory, S. (2003). Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. USA 100: 8484-8489 [Abstract] [Full Text]  
  • Rabin, S. D. P., Hauser, A. R. (2003). Pseudomonas aeruginosa ExoU, a Toxin Transported by the Type III Secretion System, Kills Saccharomyces cerevisiae. Infect. Immun. 71: 4144-4150 [Abstract] [Full Text]  
  • Jendrossek, V., Fillon, S., Belka, C., Muller, I., Puttkammer, B., Lang, F. (2003). Apoptotic Response of Chang Cells to Infection with Pseudomonas aeruginosa Strains PAK and PAO-I: Molecular Ordering of the Apoptosis Signaling Cascade and Role of Type IV Pili. Infect. Immun. 71: 2665-2673 [Abstract] [Full Text]  
  • Priebe, G. P., Meluleni, G. J., Coleman, F. T., Goldberg, J. B., Pier, G. B. (2003). Protection against Fatal Pseudomonas aeruginosa Pneumonia in Mice after Nasal Immunization with a Live, Attenuated aroA Deletion Mutant. Infect. Immun. 71: 1453-1461 [Abstract] [Full Text]  
  • Jacob, T., Lee, R. J., Engel, J. N., Machen, T. E. (2002). Modulation of Cytosolic Ca2+ Concentration in Airway Epithelial Cells by Pseudomonas aeruginosa. Infect. Immun. 70: 6399-6408 [Abstract] [Full Text]  
  • Schroeder, T. H., Lee, M. M., Yacono, P. W., Cannon, C. L., Gerceker, A. A., Golan, D. E., Pier, G. B. (2002). CFTR is a pattern recognition molecule that extracts Pseudomonas aeruginosa LPS from the outer membrane into epithelial cells and activates NF-kappa B translocation. Proc. Natl. Acad. Sci. USA 99: 6907-6912 [Abstract] [Full Text]  
  • Lyczak, J. B., Cannon, C. L., Pier, G. B. (2002). Lung Infections Associated with Cystic Fibrosis. Clin. Microbiol. Rev. 15: 194-222 [Abstract] [Full Text]  
  • Priebe, G. P., Brinig, M. M., Hatano, K., Grout, M., Coleman, F. T., Pier, G. B., Goldberg, J. B. (2002). Construction and Characterization of a Live, Attenuated aroA Deletion Mutant of Pseudomonas aeruginosa as a Candidate Intranasal Vaccine. Infect. Immun. 70: 1507-1517 [Abstract] [Full Text]  
  • Smith, R. S., Harris, S. G., Phipps, R., Iglewski, B. (2002). The Pseudomonas aeruginosa Quorum-Sensing Molecule N-(3-Oxododecanoyl)Homoserine Lactone Contributes to Virulence and Induces Inflammation In Vivo. J. Bacteriol. 184: 1132-1139 [Abstract] [Full Text]  
  • Lomholt, J. A., Poulsen, K., Kilian, M. (2001). Epidemic Population Structure of Pseudomonas aeruginosa: Evidence for a Clone That Is Pathogenic to the Eye and That Has a Distinct Combination of Virulence Factors. Infect. Immun. 69: 6284-6295 [Abstract] [Full Text]  
  • Feltman, H., Schulert, G., Khan, S., Jain, M., Peterson, L., Hauser, A. R. (2001). Prevalence of type III secretion genes in clinical and environmental isolates of Pseudomonas aeruginosa. Microbiology 147: 2659-2669 [Abstract] [Full Text]  
  • Finck-Barbancon, V., Frank, D. W. (2001). Multiple Domains Are Required for the Toxic Activity of Pseudomonas aeruginosa ExoU. J. Bacteriol. 183: 4330-4344 [Abstract] [Full Text]  
  • Schroeder, T. H., Reiniger, N., Meluleni, G., Grout, M., Coleman, F. T., Pier, G. B. (2001). Transgenic Cystic Fibrosis Mice Exhibit Reduced Early Clearance of Pseudomonas aeruginosa from the Respiratory Tract. J. Immunol. 166: 7410-7418 [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»