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B.A. Thibodeaux, A.R. Caballero, M.E. Marquart, J. Tommassen, A. Lazdunski, R.J. O'Callaghan; Virulence Contribution of Pseudomonas Aeruginosa Elastase and Alkaline Protease in a Pseudomonas Putida Model of Rabbit Keratitis . Invest. Ophthalmol. Vis. Sci. 2005;46(13):1168.
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To measure the specific virulence contributions of two Pseudomonas aeruginosa proteases, elastase B and alkaline protease, when expressed separately by Pseudomonas putida in a rabbit model of bacterial keratitis.
P. putida KT2660 was transformed with one of two plasmid constructs encoding either elastase B (lasB) or alkaline protease (aprA) along with the accessory proteins required for its secretion (aprD, aprE, aprF, and aprI). Protease expression was confirmed by zymography and Western blotting. P. putida expressing elastase B (Put:plasBT), alkaline protease (Put:pAP), or vector alone (Put:p20) was injected intrastromally (103 colony forming units [CFU] in 10 µl) into rabbit corneas (n=6). Infected corneas were graded by slit lamp examination (SLE) at 20, 24, 28, and 32 hours (h) post infection (PI). Rabbits were sacrificed at 32 h PI and the log CFU (± SEM) per cornea were determined.
SLE scores for each group at 20, 24, 28, and 32 h PI are shown in the table below.
The expression of elastase B, but not alkaline protease, in the P. putida model of keratitis significantly increased SLE scores at all time points measured. This suggests that elastase B contributes to the pathology observed during P. aeruginosa keratitis.
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