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Visualizing Macrophage Extracellular Traps Using Confocal Microscopy
Published on: October 19, 2017
Bacillus licheniformis and Bacillus subtilis, Probiotics That Induce the Formation of Macrophage Extracellular Traps
Carol M Romo-Barrera1,2, Laura E Castrillón-Rivera2, Alejandro Palma-Ramos2
1Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Ciudad de México 11340, Mexico.
Abstract:
Probiotics are considered living microorganisms that help preserve the health of the host who uses them. Bacillus are a genus of Gram-positive bacteria used as probiotics for animal and human consumption. They are currently distributed in various commercial forms. Two of the species used as probiotics are B. licheniformis and B. subtilis. Macrophages are central cells in the immune response, being fundamental in the elimination of microbial pathogens, for which they use various mechanisms, including the formation of extracellular traps (METs). There have been very few studies carried out on the participation of macrophages in response to the interaction of probiotics of the genus Bacillus with the host. In this work, we used macrophages from the J774A mouse cell line.1, and we found that they are susceptible to infection by the two Bacillus species. However, both species were eliminated as the infection progressed. Using confocal microscopy, we identified the formation of METs from the first hours of infection, which were characterized by the presence of myeloperoxidase (MPO) and citrullinated histone (Hit3Cit). Quantitative data on extracellular DNA release were also obtained; release was observed starting in the first hour of infection. The induction of METs by B. licheniformis caused a significant decrease in the colony-forming units (CFU) of Staphylococcus aureus. The induction of METS by bacteria of the Bacillus genus is a mechanism that participates in controlling the probiotic and potentially pathogenic bacteria such as S. aureus. The induction of METs to control pathogens may be a novel mechanism that could explain the beneficial effects of probiotics of the genus Bacillus.
Insights
Bacillus probiotics trigger macrophage extracellular traps (METs) to eliminate pathogens like Staphylococcus aureus. This discovery reveals a novel mechanism for probiotic benefits and pathogen control.
Area of Science:
- Immunology
- Microbiology
- Probiotics Research
Background:
- Probiotics, including Bacillus species, are beneficial microorganisms for host health.
- Macrophages are key immune cells that eliminate pathogens using mechanisms like Macrophage Extracellular Traps (METs).
- The role of macrophages in response to Bacillus probiotics is not well understood.
Purpose of the Study:
- To investigate the interaction between Bacillus licheniformis and Bacillus subtilis probiotics and macrophages.
- To determine if Bacillus species induce Macrophage Extracellular Traps (METs) formation.
- To assess the role of METs in controlling pathogenic bacteria, specifically Staphylococcus aureus.
Main Methods:
- Utilized J774A mouse macrophage cell line for in vitro studies.
- Employed confocal microscopy to visualize METs formation, characterized by myeloperoxidase (MPO) and citrullinated histone (Hit3Cit).
- Quantified extracellular DNA release and measured colony-forming units (CFU) of Staphylococcus aureus.
Main Results:
- J774A macrophages were susceptible to Bacillus infection, with bacteria being eliminated over time.
- Bacillus species induced METs formation, evidenced by extracellular DNA release starting within the first hour.
- METs induced by B. licheniformis significantly reduced Staphylococcus aureus CFU counts.
Conclusions:
- Bacillus probiotics, specifically B. licheniformis and B. subtilis, induce macrophage extracellular trap formation.
- METs play a role in controlling both probiotic Bacillus species and pathogenic bacteria like Staphylococcus aureus.
- METs induction represents a novel mechanism potentially explaining the beneficial effects of Bacillus probiotics.
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