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Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
Monitoring mRNA Expression Patterns in Macrophages in Response to Two Different Strains of Probiotics
Sang-Pil Choi1, Si-Won Park1, Seok-Jin Kang1
1Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Korea.
Abstract:
As an initial study to elucidate the molecular mechanism of how probiotics modulate macrophage activity, we monitored mRNA expression patterns in peritoneal macrophages (PMs) treated with two different strains of probiotics. After treatment with either Weissella cibaria WIKIM28 or Latilactobacillus sakei WIKIM50, total RNAs from PMs were isolated and subjected into gene chip analyses. As controls, mRNAs from vehicle (phosphate-buffered saline, PBS)-treated PMs were also subjected to gene chip analysis. Compared to vehicle (PBS)-treated PMs, WIKIM28-treated and WIKIM50-treated PMs exhibited a total of 889 and 432 differentially expressed genes with expression differences of at least 4 folds, respectively. Compared to WIKIM28-treated PMs, WIKIM50-treated PMs showed 25 up-regulated genes and 21 down-regulated genes with expression differences of more than 2 folds. Interestingly, mRNA transcripts of M2 macrophage polarization marker such as anxa1, mafb, and sepp1 were increased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. Reversely, mRNA transcripts of M1 macrophage polarization marker such as hdac9, ptgs2, and socs3 were decreased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. In agreement with these observations, mRNA expression levels of tumor necrosis factor-α and interleukin-1α were significantly reduced in WIKIM50-treated macrophages compared to those in WIKIM28-treated macrophages. These results may indicate that probiotics can be classified as two different types depending on their ability to convert macrophages into M1 or M2 polarization.
Insights
This study reveals that different probiotic strains, Weissella cibaria WIKIM28 and Latilactobacillus sakei WIKIM50, distinctly modulate macrophage polarization. Probiotics can be categorized by their influence on M1/M2 macrophage phenotypes, impacting immune responses.
Area of Science:
- Immunology
- Microbiology
- Molecular Biology
Background:
- Probiotics are known to influence host immune responses.
- The molecular mechanisms by which probiotics modulate macrophage activity remain incompletely understood.
- Macrophages play a critical role in both innate and adaptive immunity, with distinct M1 (pro-inflammatory) and M2 (anti-inflammatory/tissue repair) polarization states.
Purpose of the Study:
- To elucidate the molecular mechanisms of probiotic-mediated macrophage modulation.
- To investigate the differential effects of two specific probiotic strains, Weissella cibaria WIKIM28 and Latilactobacillus sakei WIKIM50, on macrophage polarization.
- To identify key gene expression changes associated with probiotic treatment in peritoneal macrophages.
Main Methods:
- Peritoneal macrophages (PMs) were treated with Weissella cibaria WIKIM28 or Latilactobacillus sakei WIKIM50.
- Total RNA was isolated from treated and control (vehicle-treated) PMs.
- Gene chip (microarray) analyses were performed to assess genome-wide mRNA expression patterns.
Main Results:
- Significant differential gene expression was observed in probiotic-treated PMs compared to vehicle controls.
- Latilactobacillus sakei WIKIM50 treatment increased mRNA transcripts of M2 macrophage polarization markers (e.g., anxa1, mafb, sepp1) and decreased M1 markers (e.g., hdac9, ptgs2, socs3) compared to Weissella cibaria WIKIM28.
- Tumor necrosis factor-α and interleukin-1α mRNA levels were significantly reduced in WIKIM50-treated macrophages compared to WIKIM28-treated macrophages.
Conclusions:
- Probiotic strains exhibit distinct capabilities in modulating macrophage polarization.
- Latilactobacillus sakei WIKIM50 promotes an M2-like macrophage phenotype, while Weissella cibaria WIKIM28 may induce a different response.
- These findings suggest a classification of probiotics based on their differential effects on macrophage polarization, impacting immune modulation.
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