In vivo and in vitro collagenolytic activity of Acanthamoeba castellanii

Yu-Guang He, Jerry Y Niederkorn, James P McCulley, G. L. Stewart, D. R. Meyer, R. Silvany, J. Dougherty

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Abstract

Axenic cultures of Acanthamoeba castellanii contained a collagenolytic enzyme that digested collagen shields and purified collagen in vitro. Specificity of biologic activity was determined by the addition of selected enzyme inhibitors to the assays and revealed that the parasite-conditioned medium contained both collagenase and lower concentrations of other proteolytic enzymes. However, most of the collagenolytic and pathogenic activity was directly attributable to specific collagenase. Intrastromal injection of sterile, Acanthamoeba-conditioned culture medium into naive Lewis rats produced corneal lesions clinically similar to and closely resembling those found in biopsy specimens of human patients diagnosed with acanthamoebic keratitis. Histopathologic analysis revealed moderate-to-severe neutrophil infiltration, disruption of stromal lamellae, and edema. Identical pathologic sequelae were produced by intrastromal injection of purified collagenase (25 units/ml). The pathogenicity of the soluble parasite-derived product was removed by passage over affinity columns armed with antibody specific for collagenase. These results indicated that soluble parasite-derived factors were capable of producing lesions characteristic of acanthamoebic keratitis and that the pathogenicity of these factors was either directly or indirectly attributable to specific collagenase activity.

Original languageEnglish (US)
Pages (from-to)2235-2240
Number of pages6
JournalInvestigative Ophthalmology and Visual Science
Volume31
Issue number11
StatePublished - 1990

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Acanthamoeba castellanii
Collagenases
Parasites
Keratitis
Conditioned Culture Medium
Collagen
Axenic Culture
Acanthamoeba
Injections
Neutrophil Infiltration
Enzyme Inhibitors
Virulence Factors
Virulence
Edema
Peptide Hydrolases
In Vitro Techniques
Biopsy
Antibodies
Enzymes

ASJC Scopus subject areas

  • Ophthalmology

Cite this

In vivo and in vitro collagenolytic activity of Acanthamoeba castellanii. / He, Yu-Guang; Niederkorn, Jerry Y; McCulley, James P; Stewart, G. L.; Meyer, D. R.; Silvany, R.; Dougherty, J.

In: Investigative Ophthalmology and Visual Science, Vol. 31, No. 11, 1990, p. 2235-2240.

Research output: Contribution to journalArticle

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AU - Meyer, D. R.

AU - Silvany, R.

AU - Dougherty, J.

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