Contribution of proteases and LasR to the virulence of Pseudomonas aeruginosa during corneal infections

Michael J. Preston*, Patrick C. Seed, Debbie S. Toder, Barbara H. Iglewski, Dennis E. Ohman, Jean K. Gustin, Joanna B. Goldberg, Gerald B. Pier

*Corresponding author for this work

Research output: Contribution to journalArticle

131 Scopus citations

Abstract

The roles of the Pseudomonas aeruginosa proteases LasB (elastase) and LasA and the transcriptional activator LasR, which regulates the expression of these proteases, were evaluated in a murine model of P. aeruginosa corneal infection. In scarified corneas, P. aeruginosa PAO-A1 (LasA negative) or PAO- B1A1 (LasB and LasA negative) at a dose of 108 CFU per eye caused very mild or no disease following infection; however, the defect in PAO-A1 could not be complemented by supplying a functional copy of lasA either on a plasmid or inserted into the chromosome. In contrast, PAO-B1 (LasB negative) colonized the cornea and caused disease equal in severity to disease caused by the parental strain, PAO1-I. Although LasR is a known regulator of lasA expression, PAO-R1, a lasR-negative derivative of PAO1-I, was as virulent as the parental strain during corneal infection. When transcriptional fusion plasmids were used to quantify the expression of the lasB and lasA genes in P. aeruginosa PAO1-I and PAO-R1, the lasB::lacZ fusion in PAO-R1 showed only 3.5% as much activity as it did in PAO1-I, while the activity of the lasA::lacZ fusion in PAO-R1 was 27.8% of that in PAO1-I. Coadministration of 5 μg of purified LasA protease with PAO-A1 did not reconstitute a wild-type infection. This treatment produced an acute toxic reaction leading to prolonged eyelid closure without inflammatory destruction of the cornea that was similar to that observed when LasA was administered alone. These results indicate that insertional inactivation of lasA renders P. aeruginosa avirulent in a murine model of keratitis and that neither LasR nor elastase production is required for the establishment and maintenance of corneal infection. However, the lack of virulence of the LasA-deficient strains cannot be ascribed with certainty to the deficiency of LasA from the available data.

Original languageEnglish (US)
Pages (from-to)3086-3090
Number of pages5
JournalInfection and immunity
Volume65
Issue number8
DOIs
StatePublished - 1997

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

Fingerprint Dive into the research topics of 'Contribution of proteases and LasR to the virulence of Pseudomonas aeruginosa during corneal infections'. Together they form a unique fingerprint.

  • Cite this

    Preston, M. J., Seed, P. C., Toder, D. S., Iglewski, B. H., Ohman, D. E., Gustin, J. K., Goldberg, J. B., & Pier, G. B. (1997). Contribution of proteases and LasR to the virulence of Pseudomonas aeruginosa during corneal infections. Infection and immunity, 65(8), 3086-3090. https://doi.org/10.1128/iai.65.8.3086-3090.1997