June 1995
Volume 36, Issue 7
Free
Articles  |   June 1995
In vivo bacterial protease production during Pseudomonas aeruginosa corneal infection.
Author Affiliations
  • K A Kernacki
    Department of Immunology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
  • J A Hobden
    Department of Immunology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
  • L D Hazlett
    Department of Immunology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
  • R Fridman
    Department of Immunology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
  • R S Berk
    Department of Immunology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Investigative Ophthalmology & Visual Science June 1995, Vol.36, 1371-1378. doi:
  • Views
  • PDF
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      K A Kernacki, J A Hobden, L D Hazlett, R Fridman, R S Berk; In vivo bacterial protease production during Pseudomonas aeruginosa corneal infection.. Invest. Ophthalmol. Vis. Sci. 1995;36(7):1371-1378.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

PURPOSE: To establish if active pseudomonal proteases are present in vivo during corneal infection with Pseudomonas aeruginosa and to determine if the mouse strains used in these and previous studies have the ability to mount a nonocular antibody response to the purified proteases because antibodies to the bacterial proteases were not detected previously during in vivo ocular infection. METHODS: At certain times after corneal infection with P. aeruginosa, corneas were harvested and supernatants from the corneal homogenates were analyzed for proteolytic activity by zymography and immunoreactivity by immunoblotting. The efficiency of the extraction procedures used in these studies was determined by incubating uninfected corneal homogenates with the purified proteases. The resultant supernatants were analyzed for alkaline protease and elastase activity. Additionally, mice were immunized intraperitoneally with the purified proteases with and without adjuvant to determine if the animals could mount a nonocular antibody response. RESULTS: Corneas infected with P. aeruginosa demonstrated the presence of alkaline protease, but not elastase, by the two methods examined. The kinetics of the in vivo alkaline protease response closely parallels previously reported bacterial clearance studies in that peak alkaline protease activity was detected in corneal tissue when peak bacterial numbers also were observed in the eye, and it was absent when the eyes were sterile or nearly sterile. In addition, C57BL/6J mice were capable of mounting a nonocular antibody response to microgram quantities of both proteases only in the presence of adjuvant. CONCLUSIONS: In the model described, enzymatically active alkaline protease, but not elastase, was demonstrated in corneal tissues during in vivo infection. Concentrations of these proteases were much lower than those required to stimulate an antibody response.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×