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Isolation of Murine Peritoneal Macrophages to Carry Out Gene Expression Analysis Upon Toll-like Receptors Stimulation
Published on: April 29, 2015
Protein expression changes induced in murine peritoneal macrophages by Group B Streptococcus
Federica Susta1, Davide Chiasserini, Katia Fettucciari
1Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Perugia, Italy.
Proteomics
|March 26, 2010
Summary
Group B Streptococcus (GBS) infection alters protein expression in murine macrophages, impacting phagocytosis, stress responses, and energy metabolism. Key enzymes for reactive oxygen species and glycolysis were notably downregulated.
Area of Science:
- Immunology
- Microbiology
- Proteomics
Background:
- Murine macrophages are crucial in host defense against bacterial infections.
- Group B Streptococcus (GBS) is a significant pathogen, particularly in neonates.
- Understanding host-pathogen interactions at the protein level is vital for developing therapeutic strategies.
Purpose of the Study:
- To investigate the proteomic changes in murine macrophages upon infection with GBS.
- To identify specific proteins and pathways affected by GBS, particularly those related to immune function and metabolism.
Main Methods:
- Thioglycolate-elicited peritoneal murine macrophages were infected with live or heat-inactivated GBS.
- Proteins were separated using two-dimensional gel electrophoresis (2-DE).
- Differentially expressed proteins were identified using tandem mass spectrometry (MS/MS analysis).
Main Results:
- GBS infection significantly altered the expression of proteins involved in phagocytic modulation, stress response, and cell death.
- Expression of enzymes critical for reactive oxygen species production was decreased in infected macrophages.
- Metabolic enzymes, including glycolytic and pentose-cycle enzymes, were predominantly downregulated in macrophages infected with live GBS.
- GBS infection impacted the expression of enzymes involved in ATP synthesis and adenine nucleotide metabolism.
Conclusions:
- GBS infection induces complex proteomic alterations in macrophages, affecting critical cellular functions.
- The downregulation of key metabolic and reactive oxygen species-producing enzymes suggests a mechanism by which GBS evades host defenses.
- These findings provide insights into macrophage dysfunction during GBS infection and potential therapeutic targets.
