Detection of rat Pneumocystis carinii proteinases and elastase and antipneumocystis activity of proteinase inhibitors in vitro.
Proteinases play an important role in survival of microorganisms and in pathogenicity of diseases. By using a modified SDS-gelatin-polyacrylamide gel system, proteinases of rat-P.carinii were detected as bands of proteolytic digestion after electrophoresis. P.carinii organisms obtained from dexamethasone immunosuppressed transtracheally infected rats were cultured in spinner flask suspension cultures to minimize host cell contamination. At pH 8.3, seven Pc-specific proteolytic bands were detected in three clusters of different molecular weights clearly different from host cell patterns. By using a range of pH, various preparations of organisms and both infected and uninfected culture media, proteolytic activities have been partially characterized. Elastase secretion has been assessed based on elastin digestion model. Proteinase inhibitors have been tested for their ability to inhibit P.carinii growth in HEL299 short-term monolayer cultures. Results indicate that proteolytic activities are involved in the proliferation of microorganisms since leupeptin exerted in vitro antipneumocystis activity while aprotinin enhanced P.carinii growth.
Atzori
C
C
II Div. Infect. Dis., L. Sacco Hospital, Milan, Italy.
Mainini
A
A
Agostoni
F
F
Angeli
E
E
Bartlett
M
M
Bruno
A
A
Scaglia
M
M
Cargnel
A
A
D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D004549:Elastin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000032:analysis*; D005260:Female; D005656:Fungal Proteins / Q000032:analysis* / Q000037:antagonists & inhibitors; D005780:Gelatin / Q000378:metabolism; D010196:Pancreatic Elastase / Q000032:analysis* / Q000037:antagonists & inhibitors; D011010:Pneumocystis / Q000187:drug effects / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity
D000935:Antifungal Agents; D005656:Fungal Proteins; D011480:Protease Inhibitors; D005780:Gelatin; D004549:Elastin; D010450:Endopeptidases; D010196:Pancreatic Elastase