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Updated: Apr 23, 2026
Establishing a Porcine Ex Vivo Cornea Model for Studying Drug Treatments against Bacterial Keratitis
Published on: May 12, 2020
Proteomic analysis of keratitis-associated Pseudomonas aeruginosa
Abby Sewell1, Jeffrey Dunmire1, Michael Wehmann1
1Summa Health System, Akron, OH.
Purpose:
To compare the proteomic profile of a clinical isolate of Pseudomonas aeruginosa (P. aeruginosa) obtained from an infected cornea of a contact lens wearer and the laboratory strain P. aeruginosa ATCC 10145.
Methods:
Antibiotic sensitivity, motility, biofilm formation, and virulence tests were performed using standard methods. Whole protein lysates were analyzed with liquid chromatography/ tandem mass spectrometry (LC-MS/MS) in triplicate, and relative protein abundances were determined with spectral counting. The G test followed by a post hoc Holm-Sidak adjustment was used for the statistical analyses to determine significance in the differential expression of proteins between the two strains.
Results:
A total of 687 proteins were detected. One-hundred thirty-three (133) proteins were significantly different between the two strains. Among these, 13 were upregulated, and 16 were downregulated in the clinical strain compared to ATCC 10145, whereas 57 were detected only in the clinical strain. The upregulated proteins are associated with virulence and pathogenicity.
Conclusions:
Proteins detected at higher levels in the clinical strain of P. aeruginosa were proteins known to be virulence factors. These results confirm that the keratitis-associated P. aeruginosa strain is pathogenic and expresses a higher number of virulence factors compared to the laboratory strain ATCC 10145. Identification of the protein profile of the corneal strain of P. aeruginosa in this study will aid in elucidating novel intervention strategies for reducing the burden of P. aeruginosa infection in keratitis.
Insights
A clinical Pseudomonas aeruginosa strain from a contact lens wearer
Area of Science:
- Microbiology
- Proteomics
- Ophthalmology
Background:
- Pseudomonas aeruginosa (P. aeruginosa) is a common cause of microbial keratitis.
- Contact lens wear is a significant risk factor for P. aeruginosa keratitis.
- Understanding the molecular differences between clinical and laboratory strains is crucial for developing targeted therapies.
Purpose of the Study:
- To compare the proteomic profiles of a clinical P. aeruginosa isolate from a contact lens wearer's cornea with a laboratory strain (ATCC 10145).
- To identify differentially expressed proteins and their potential roles in pathogenicity.
Main Methods:
- Standard microbiological tests (antibiotic sensitivity, motility, biofilm formation, virulence).
- Liquid chromatography/tandem mass spectrometry (LC-MS/MS) for whole protein lysate analysis.
- Statistical analysis using the G test with Holm-Sidak adjustment for differential protein expression.
Main Results:
- A total of 687 proteins were detected.
- 133 proteins showed significant differences between the clinical and laboratory strains.
- 57 proteins were unique to the clinical strain, and upregulated proteins were associated with virulence and pathogenicity.
Conclusions:
- The clinical P. aeruginosa strain exhibits a higher expression of virulence factors compared to the laboratory strain.
- This confirms the pathogenicity of the keratitis-associated strain.
- Identifying the protein profile of the corneal strain can guide novel intervention strategies for P. aeruginosa keratitis.
