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Histamine derived from probiotic Lactobacillus reuteri suppresses TNF via modulation of PKA and ERK signaling.

Thomas CM, Hong T, van Pijkeren JP, Hemarajata P, Trinh DV, Hu W, Britton RA, Kalkum M, Versalovic J - PLoS ONE (2012)

Bottom Line: Previously, immunomodulatory factors secreted by L. reuteri ATCC PTA 6475 were unknown.A combined metabolomics and bacterial genetics strategy was utilized to identify small compound(s) produced by L. reuteri that were TNF-inhibitory.The identification of bacterial bioactive metabolites and their corresponding mechanisms of action with respect to immunomodulation may lead to improved anti-inflammatory strategies for chronic immune-mediated diseases.

View Article: PubMed Central - PubMed

Affiliation: Interdepartmental Program in Cell and Molecular Biology, Baylor College of Medicine, Houston, Texas, United States of America.

ABSTRACT
Beneficial microbes and probiotic species, such as Lactobacillus reuteri, produce biologically active compounds that can modulate host mucosal immunity. Previously, immunomodulatory factors secreted by L. reuteri ATCC PTA 6475 were unknown. A combined metabolomics and bacterial genetics strategy was utilized to identify small compound(s) produced by L. reuteri that were TNF-inhibitory. Hydrophilic interaction liquid chromatography-high performance liquid chromatography (HILIC-HPLC) separation isolated TNF-inhibitory compounds, and HILIC-HPLC fraction composition was determined by NMR and mass spectrometry analyses. Histamine was identified and quantified in TNF-inhibitory HILIC-HPLC fractions. Histamine is produced from L-histidine via histidine decarboxylase by some fermentative bacteria including lactobacilli. Targeted mutagenesis of each gene present in the histidine decarboxylase gene cluster in L. reuteri 6475 demonstrated the involvement of histidine decarboxylase pyruvoyl type A (hdcA), histidine/histamine antiporter (hdcP), and hdcB in production of the TNF-inhibitory factor. The mechanism of TNF inhibition by L. reuteri-derived histamine was investigated using Toll-like receptor 2 (TLR2)-activated human monocytoid cells. Bacterial histamine suppressed TNF production via activation of the H(2) receptor. Histamine from L. reuteri 6475 stimulated increased levels of cAMP, which inhibited downstream MEK/ERK MAPK signaling via protein kinase A (PKA) and resulted in suppression of TNF production by transcriptional regulation. In summary, a component of the gut microbiome, L. reuteri, is able to convert a dietary component, L-histidine, into an immunoregulatory signal, histamine, which suppresses pro-inflammatory TNF production. The identification of bacterial bioactive metabolites and their corresponding mechanisms of action with respect to immunomodulation may lead to improved anti-inflammatory strategies for chronic immune-mediated diseases.

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Identification of histamine in TNF-inhibitory fractions of L. reuteri 6475 by NMR.1H NMR analysis demonstrated differences in the composition of HILIC-HPLC fractions from L. reuteri 6475. A. Top and bottom spectra show the 1D 1H NMR of TNF-inhibitory fraction B3 and fraction B4, respectively. The assigned peaks of phenylalanine (Phe), tryptophan (Trp), and histamine from fraction B3 are labeled on the top spectrum. Two complementary 2D NMR techniques were used to identify compounds unique to TNF-inhibitory fraction B3. B. Aromatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. C. Aliphatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. D. The TOCSY spectrum further identified the spin system of Phe, Trp, and histamine unique to TNF-inhibitory fraction B3.
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pone-0031951-g001: Identification of histamine in TNF-inhibitory fractions of L. reuteri 6475 by NMR.1H NMR analysis demonstrated differences in the composition of HILIC-HPLC fractions from L. reuteri 6475. A. Top and bottom spectra show the 1D 1H NMR of TNF-inhibitory fraction B3 and fraction B4, respectively. The assigned peaks of phenylalanine (Phe), tryptophan (Trp), and histamine from fraction B3 are labeled on the top spectrum. Two complementary 2D NMR techniques were used to identify compounds unique to TNF-inhibitory fraction B3. B. Aromatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. C. Aliphatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. D. The TOCSY spectrum further identified the spin system of Phe, Trp, and histamine unique to TNF-inhibitory fraction B3.

Mentions: The TNF-inhibitory HILIC-HPLC fraction B3 was analyzed by one-dimensional (1D) 1H NMR and compared to the neighboring non-TNF-inhibitory fraction B4. A unique series of peaks with a chemical shift between 7.0–8.0 ppm, which indicate the presence of aromatic or heterocyclic compounds, were observed in fraction B3 (Figure 1A, top spectrum) but not in fraction B4 (Figure 1A, bottom spectrum). To identify the compounds yielding these peaks, two-dimensional (2D) 1H-13C-heteronuclear single quantum coherence (HSQC) data were acquired with fraction B3, and compounds were identified using MetaboMiner software [27]. The compounds present in fraction B3 were tryptophan (Trp), phenylalanine (Phe), histamine, and one unknown peak (Figure 1B and 1C). All observable 1H-13C cross peaks of these three compounds are labeled in Figure 1B and 1C, which were identical to those compounds listed in the database of MetaboMiner. Compound identification was further confirmed using an additional 2D NMR method, total correlation spectroscopy (TOCSY), as shown in Figure 1D. Tryptophan and phenylalanine were components of the defined bacterial growth medium. Histidine, but not histamine, was part of the bacterial growth medium, suggesting that histidine may be converted to histamine by L. reuteri.


Histamine derived from probiotic Lactobacillus reuteri suppresses TNF via modulation of PKA and ERK signaling.

Thomas CM, Hong T, van Pijkeren JP, Hemarajata P, Trinh DV, Hu W, Britton RA, Kalkum M, Versalovic J - PLoS ONE (2012)

Identification of histamine in TNF-inhibitory fractions of L. reuteri 6475 by NMR.1H NMR analysis demonstrated differences in the composition of HILIC-HPLC fractions from L. reuteri 6475. A. Top and bottom spectra show the 1D 1H NMR of TNF-inhibitory fraction B3 and fraction B4, respectively. The assigned peaks of phenylalanine (Phe), tryptophan (Trp), and histamine from fraction B3 are labeled on the top spectrum. Two complementary 2D NMR techniques were used to identify compounds unique to TNF-inhibitory fraction B3. B. Aromatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. C. Aliphatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. D. The TOCSY spectrum further identified the spin system of Phe, Trp, and histamine unique to TNF-inhibitory fraction B3.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3285189&req=5

pone-0031951-g001: Identification of histamine in TNF-inhibitory fractions of L. reuteri 6475 by NMR.1H NMR analysis demonstrated differences in the composition of HILIC-HPLC fractions from L. reuteri 6475. A. Top and bottom spectra show the 1D 1H NMR of TNF-inhibitory fraction B3 and fraction B4, respectively. The assigned peaks of phenylalanine (Phe), tryptophan (Trp), and histamine from fraction B3 are labeled on the top spectrum. Two complementary 2D NMR techniques were used to identify compounds unique to TNF-inhibitory fraction B3. B. Aromatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. C. Aliphatic peaks of 1H-13C-HSQC from Phe, Trp, and histamine. D. The TOCSY spectrum further identified the spin system of Phe, Trp, and histamine unique to TNF-inhibitory fraction B3.
Mentions: The TNF-inhibitory HILIC-HPLC fraction B3 was analyzed by one-dimensional (1D) 1H NMR and compared to the neighboring non-TNF-inhibitory fraction B4. A unique series of peaks with a chemical shift between 7.0–8.0 ppm, which indicate the presence of aromatic or heterocyclic compounds, were observed in fraction B3 (Figure 1A, top spectrum) but not in fraction B4 (Figure 1A, bottom spectrum). To identify the compounds yielding these peaks, two-dimensional (2D) 1H-13C-heteronuclear single quantum coherence (HSQC) data were acquired with fraction B3, and compounds were identified using MetaboMiner software [27]. The compounds present in fraction B3 were tryptophan (Trp), phenylalanine (Phe), histamine, and one unknown peak (Figure 1B and 1C). All observable 1H-13C cross peaks of these three compounds are labeled in Figure 1B and 1C, which were identical to those compounds listed in the database of MetaboMiner. Compound identification was further confirmed using an additional 2D NMR method, total correlation spectroscopy (TOCSY), as shown in Figure 1D. Tryptophan and phenylalanine were components of the defined bacterial growth medium. Histidine, but not histamine, was part of the bacterial growth medium, suggesting that histidine may be converted to histamine by L. reuteri.

Bottom Line: Previously, immunomodulatory factors secreted by L. reuteri ATCC PTA 6475 were unknown.A combined metabolomics and bacterial genetics strategy was utilized to identify small compound(s) produced by L. reuteri that were TNF-inhibitory.The identification of bacterial bioactive metabolites and their corresponding mechanisms of action with respect to immunomodulation may lead to improved anti-inflammatory strategies for chronic immune-mediated diseases.

View Article: PubMed Central - PubMed

Affiliation: Interdepartmental Program in Cell and Molecular Biology, Baylor College of Medicine, Houston, Texas, United States of America.

ABSTRACT
Beneficial microbes and probiotic species, such as Lactobacillus reuteri, produce biologically active compounds that can modulate host mucosal immunity. Previously, immunomodulatory factors secreted by L. reuteri ATCC PTA 6475 were unknown. A combined metabolomics and bacterial genetics strategy was utilized to identify small compound(s) produced by L. reuteri that were TNF-inhibitory. Hydrophilic interaction liquid chromatography-high performance liquid chromatography (HILIC-HPLC) separation isolated TNF-inhibitory compounds, and HILIC-HPLC fraction composition was determined by NMR and mass spectrometry analyses. Histamine was identified and quantified in TNF-inhibitory HILIC-HPLC fractions. Histamine is produced from L-histidine via histidine decarboxylase by some fermentative bacteria including lactobacilli. Targeted mutagenesis of each gene present in the histidine decarboxylase gene cluster in L. reuteri 6475 demonstrated the involvement of histidine decarboxylase pyruvoyl type A (hdcA), histidine/histamine antiporter (hdcP), and hdcB in production of the TNF-inhibitory factor. The mechanism of TNF inhibition by L. reuteri-derived histamine was investigated using Toll-like receptor 2 (TLR2)-activated human monocytoid cells. Bacterial histamine suppressed TNF production via activation of the H(2) receptor. Histamine from L. reuteri 6475 stimulated increased levels of cAMP, which inhibited downstream MEK/ERK MAPK signaling via protein kinase A (PKA) and resulted in suppression of TNF production by transcriptional regulation. In summary, a component of the gut microbiome, L. reuteri, is able to convert a dietary component, L-histidine, into an immunoregulatory signal, histamine, which suppresses pro-inflammatory TNF production. The identification of bacterial bioactive metabolites and their corresponding mechanisms of action with respect to immunomodulation may lead to improved anti-inflammatory strategies for chronic immune-mediated diseases.

Show MeSH
Related in: MedlinePlus