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Release of lysosomal enzymes in Candida albicans phagocytosis by rat peritoneal macrophages
Abstract:
The present paper reports the in vitro release of lysosomal enzymes in the supernatant of cultures of rat peritoneal macrophages, with the addition of Candida albicans cells. Macrophages were taken from the rat peritoneal cavity 72 hr after non-specific activation with Brain-Heart-Infusion (B.H.I.) broth containing 10% proteose-peptone No. 3. They were then cultured in Parker medium No. 199 (TC 199). After 24 hr a suspension of Candida albicans cells, in a determined concentration, was added to the peritoneal macrophage cultures. At that time, and during pre-determined periods, the following enzymes in the culture supernatants were studied using colorimetric methods: beta-glucuronidase, beta-galactosidase and acid phosphatase. It is concluded that, under identical conditions, the release of beta-galactosidase and acid phosphatase is higher than for beta-glucuronidase. The release rate of all three enzymes is the highest at a 6 hr incubation period, after which, a gradual decrease leads to the rate down to 50% at 24 hr.
Insights
This study investigated lysosomal enzyme release from rat macrophages exposed to Candida albicans. Beta-galactosidase and acid phosphatase showed higher release than beta-glucuronidase, peaking at 6 hours.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Rat peritoneal macrophages are key immune cells involved in pathogen clearance.
- Candida albicans is a common fungal pathogen that can be recognized and phagocytosed by macrophages.
- Lysosomal enzymes are crucial for degrading engulfed microbial components.
Purpose of the Study:
- To quantify the in vitro release of specific lysosomal enzymes from activated rat macrophages upon challenge with Candida albicans.
- To compare the release kinetics of beta-glucuronidase, beta-galactosidase, and acid phosphatase.
Main Methods:
- Primary cultures of rat peritoneal macrophages were activated using Brain-Heart-Infusion (B.H.I.) broth.
- Macrophages were cultured in Parker medium No. 199 (TC 199).
- Candida albicans cells were added to macrophage cultures, and enzyme release into the supernatant was measured using colorimetric assays at various time points.
Main Results:
- Beta-galactosidase and acid phosphatase exhibited significantly higher release rates compared to beta-glucuronidase under identical experimental conditions.
- The highest release rate for all three lysosomal enzymes was observed at the 6-hour incubation mark.
- Enzyme release rates gradually decreased after 6 hours, reaching approximately 50% of the peak levels by 24 hours.
Conclusions:
- The study demonstrates differential release patterns of lysosomal enzymes from activated macrophages in response to Candida albicans.
- Beta-galactosidase and acid phosphatase are released more readily than beta-glucuronidase by macrophages challenged with this fungus.
- Macrophage lysosomal enzyme release kinetics show a peak activity around 6 hours post-challenge, followed by a decline.