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Study of Phagolysosome Biogenesis in Live Macrophages
Published on: March 10, 2014
Protein expression changes during phagocytosis influenced by low-frequency electromagnetic field exposure
Piotr Piszczek1, Karolina Wójcik-Piotrowicz2, Piotr Guzdek3
1Department of Pathophysiology, Faculty of Medicine, Jagiellonian University Medical College, Czysta street 18, 31-121 Cracow, Poland.
Abstract:
The aim of our studies was to determine the influence of a low-frequency electromagnetic field (EMF) on the phagocytosis of latex beads (LBs) and the expression level of proteins/genes in the human monocytic macrophage Mono Mac 6 (MM6) cell line in in vitro conditions. Before phagocytosis assay cells were pre-stimulated with infectious agents such as lipopolysaccharide (LPS), Staphylococcal enterotoxin B (SEB), or the proliferatory agent phytohaemagglutinin (PHA), and then exposed to EMF (30 mT, 7 Hz, 3 h). The expression of cytoplasmic proteins like iPLA, cPLA, iNOS, NLR3/4, and Hsp70 involved in the immune response pathways to phagocytosed particles were evaluated with the usage of the Western blot analysis. mRNA encoding the iNOS protein was detected by reverse transcription PCR method. The most meaningful changes were observed for PLA2 and NLC4 proteins level and between iNOS protein expression and mRNA encoding iNOS protein amount. The EMF exposure exerted the strongest effect on iNOS encoding mRNA in cells pre-stimulated with LPS or SEB and phagocytosing LBs. The influence of EMF on phagocytosis was experimentally proved for the first time and there is a need for further investigations in term of the usage of EMF as a prospect, supportive therapy.
Insights
Low-frequency electromagnetic fields (EMF) significantly altered immune gene expression in macrophages. EMF exposure influenced iNOS mRNA levels, particularly after stimulation with LPS or SEB, suggesting potential therapeutic applications.
Area of Science:
- Immunology
- Biophysics
Background:
- Macrophages play a crucial role in the innate immune system, engulfing pathogens and cellular debris through phagocytosis.
- Immune responses involve complex signaling pathways regulated by protein and gene expression.
- Low-frequency electromagnetic fields (EMF) are being explored for their potential biological effects.
Purpose of the Study:
- To investigate the impact of low-frequency EMF on macrophage phagocytosis and immune-related protein/gene expression.
- To assess changes in specific proteins (iPLA, cPLA, iNOS, NLR3/4, Hsp70) and iNOS mRNA levels in human monocytic macrophages (MM6) after EMF exposure.
Main Methods:
- Human monocytic macrophage (MM6) cell line was used for in vitro studies.
- Cells were pre-stimulated with lipopolysaccharide (LPS), Staphylococcal enterotoxin B (SEB), or phytohaemagglutinin (PHA).
- Phagocytosis assay with latex beads (LBs) was performed, followed by EMF exposure (30 mT, 7 Hz, 3 h).
- Protein expression was analyzed using Western blot; mRNA levels were determined by reverse transcription PCR.
Main Results:
- EMF exposure modulated the expression of several immune-related proteins, with notable changes in PLA2 and NLC4 levels.
- Significant alterations were observed in the expression of inducible nitric oxide synthase (iNOS) protein and its corresponding mRNA.
- The strongest effect of EMF on iNOS mRNA was seen in cells pre-stimulated with LPS or SEB and undergoing phagocytosis of LBs.
Conclusions:
- This study provides the first experimental evidence of EMF influencing phagocytosis.
- EMF exposure significantly impacts iNOS gene expression in macrophages, particularly under specific immune stimulation conditions.
- Further research is warranted to explore the potential of EMF as a supportive therapy in immune modulation.
