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In Vitro Comparison of the Effects of Probiotic, Commensal and Pathogenic Strains on Macrophage Polarization

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Abstract

Macrophages are important with respect to both innate and adaptive immune responses and are known to differentiate into pro-inflammatory M1- or anti-inflammatory M2-phenotypes following activation. In order to study how different bacteria affect macrophage polarization, we exposed murine RAW 264.7 macrophages to sixteen different strains representing probiotic strains, pathogens, commensals and strains of food origin. Increased inducible nitric oxide synthase (iNOS) or arginase-1 gene expression indicates M1 or M2 polarization, respectively, and was quantified by qRT-PCR. Strains of Escherichia and Salmonella elevated iNOS expression more so than strains of Enterococcus, Lactobacillus and Lactococcus, indicating that Gram-negative strains are more potent M1 inducers. However, strain-specific responses were observed. For instance, Escherichia coli Nissle 1917 was a poor inducer of iNOS gene expression compared to the other E. coli strains, while Enterococcus faecalis Symbioflor-1 was more potent in this respect compared to all the eleven Gram-positive strains tested. Macrophage polarization was further characterized by quantifying secreted pro- and anti-inflammatory cytokines. Exposure to the pathogen E. coli 042 produced a cytokine profile indicating M1 differentiation, which is in accordance with the PCR data. However, exposure to most strains resulted in either high or low secretion levels of all cytokines tested, rather than a clear M1 or M2 profile. In general, the Gram-negative strains induced high levels of cytokine secretion compared to the Gram-positive strains. Interestingly, strains of human origin had a higher impact on macrophages compared to strains of food origin.

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Acknowledgments

This work was supported by the Ostfold Hospital Trust. Post-doctor Alasdair Mckenzie was most helpful in reviewing the manuscript.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Faculty of Engineering, Ostfold University College, 1757, Halden, Norway

    Trine Eker Christoffersen

  2. Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1432, Ås, Norway

    Trine Eker Christoffersen, Lene Therese Olsen Hult, Katarzyna Kuczkowska, Kim Marius Moe, Siv Skeie, Tor Lea & Charlotte Ramstad Kleiveland

  3. Ostfold Hospital Trust, 1603, Fredrikstad, Norway

    Lene Therese Olsen Hult & Charlotte Ramstad Kleiveland

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  1. Trine Eker Christoffersen
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Correspondence to Trine Eker Christoffersen.

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Trine Eker Christoffersen and Lene Therese Olsen Hult have contributed equally to this work.

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Christoffersen, T.E., Hult, L.T.O., Kuczkowska, K. et al. In Vitro Comparison of the Effects of Probiotic, Commensal and Pathogenic Strains on Macrophage Polarization. Probiotics & Antimicro. Prot. 6, 1–10 (2014). https://doi.org/10.1007/s12602-013-9152-0

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