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Inhibition of rat alveolar macrophage phagocytic function by a Pseudomonas cepacia lipase
D C Straus1, M K Lonon, J C Hutson
1Department of Microbiology, Texas Tech University Health Sciences Center, Lubbock 79430.
Abstract:
The effects of purified Pseudomonas cepacia lipase on rat pulmonary alveolar function and morphology were examined. Lipase (2.5-20 micrograms/ml) adversely effected the phagocytic function of rat pulmonary alveolar macrophages in a dose-dependent manner. The lipase itself was not directly cytotoxic to these cells. Alveolar macrophages, in the absence of lipase, phagocytosed c. 35% of a given population of opsonised P. cepacia in 30 min when the ratio of bacteria:phagocyte was 10:1. Phagocytosis of P. cepacia by rat pulmonary alveolar macrophages was significantly reduced when the cells were either pre-incubated with the lipase or when phagocytosis occurred in the presence of the lipase. This was confirmed by transmission electronmicroscopy. These functional changes were associated with marked alterations of the macrophage morphology. Scanning electronmicroscopy showed that macrophages exposed to the P. cepacia lipase had fewer specialised surface structures and did not spread on plastic surfaces as well as untreated macrophages. The effects of the lipase were lost after heat inactivation, which indicates that the effects of the P. cepacia lipase were due to its enzymic activity. These results suggest that, if sufficient quantities of the enzyme are produced in vivo, lipase may be an important virulence factor for P. cepacia, allowing the organism to evade phagocytic cells.
Insights
Pseudomonas cepacia lipase impairs rat pulmonary alveolar macrophage function and alters cell structure. This enzyme activity may help the bacteria evade immune cells, acting as a virulence factor.
Area of Science:
- Microbiology
- Immunology
- Enzymology
Background:
- Pseudomonas cepacia is an opportunistic pathogen.
- Bacterial lipases can play roles in virulence.
- Pulmonary alveolar macrophages are key immune cells in the lungs.
Purpose of the Study:
- To investigate the impact of Pseudomonas cepacia lipase on rat pulmonary alveolar macrophage function and morphology.
- To determine if lipase activity contributes to bacterial evasion of phagocytosis.
Main Methods:
- Exposure of rat pulmonary alveolar macrophages to purified Pseudomonas cepacia lipase.
- Assessment of phagocytic activity using opsonised P. cepacia.
- Microscopic analysis (transmission and scanning electron microscopy) of macrophage morphology.
- Heat inactivation of lipase to confirm enzyme-dependent effects.
Main Results:
- Lipase reduced macrophage phagocytosis of P. cepacia in a dose-dependent manner.
- Lipase caused significant alterations in macrophage morphology, including reduced surface structures.
- Effects were abolished by heat inactivation, confirming enzyme activity.
- Lipase was not directly cytotoxic to macrophages.
Conclusions:
- Pseudomonas cepacia lipase inhibits the phagocytic function of pulmonary alveolar macrophages.
- Lipase-induced morphological changes may impair macrophage effectiveness.
- Enzymatic activity of lipase is crucial for these effects.
- Lipase is a potential virulence factor for P. cepacia, aiding immune evasion.