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Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages
Published on: November 3, 2014
[Inhibition of functional activity of macrophages in vitro by dimer RNase Bacillus intermedius]
Abstract:
The influence of cross-linked by dimethylsuberimidate dimeric RNAse from Bacillus intermedius on peritoneal rat macrophages has been investigated in vitro. It has been shown that dimeric RNase with concentrations of 0.5-40.0 mg/ml decreases the functional activities of macrophages. This is manifested in the inhibition of the phagocyte function of macrophages and suppression of the fusion of phagosomes with lysosomes. The change in the cytoplasmatic membrane surface structure induced by the dimers, which is stronger than that induced by monomers, has been demonstrated using atomic force microscopy. The role of membrane properties modification in the inhibition effect of RNase dimers on the functional activities of macrophages is discussed.
Insights
Dimeric RNAse from Bacillus intermedius inhibits rat macrophage functions, including phagocytosis and phagosome-lysosome fusion. This enzyme alters cell membrane structure more significantly than monomers.
Area of Science:
- Immunology
- Biochemistry
- Enzymology
Background:
- Macrophages are crucial immune cells involved in phagocytosis and pathogen clearance.
- Bacillus intermedius ribonuclease (RNase) is an enzyme with potential immunomodulatory effects.
- Dimeric forms of enzymes can exhibit distinct biological activities compared to monomers.
Purpose of the Study:
- To investigate the in vitro effects of dimeric RNAse from Bacillus intermedius on rat peritoneal macrophages.
- To determine the impact of dimeric RNase on macrophage functional activities, including phagocytosis and lysosomal fusion.
- To explore the structural changes induced by dimeric RNase on macrophage cell membranes.
Main Methods:
- In vitro incubation of rat peritoneal macrophages with varying concentrations of dimeric RNase.
- Assessment of macrophage functional activities: phagocyte function and phagosome-lysosome fusion.
- Atomic force microscopy (AFM) to analyze changes in cell membrane surface structure.
Main Results:
- Dimeric RNase significantly decreased macrophage functional activities at concentrations of 0.5-40.0 mg/ml.
- Inhibition of both phagocyte function and the fusion of phagosomes with lysosomes was observed.
- Atomic force microscopy revealed that dimeric RNase induced more pronounced changes in the cytoplasmatic membrane surface structure compared to monomeric RNase.
Conclusions:
- Dimeric RNAse from Bacillus intermedius exerts an inhibitory effect on rat macrophage functions.
- The observed inhibition is linked to the suppression of phagocytosis and impaired phagosome-lysosome fusion.
- Alterations in macrophage membrane properties induced by dimeric RNase play a role in its inhibitory effects.
