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Characterization of the anti-inflammatory Lactobacillus reuteri BM36301 and its probiotic benefits on aged mice.

Lee J, Yang W, Hostetler A, Schultz N, Suckow MA, Stewart KL, Kim DD, Kim HS - BMC Microbiol. (2016)

Bottom Line: On the other hand, treatment of THP-1 directly with live bacterial cells identified a group of pro-inflammatory LAB, which stimulated significant production of TNF-α.Furthermore, while males treated with pro-inflammatory BM36304 developed higher serum levels of TNF-α and insulin, in contrast females did not experience such effects from this bacteria strain.It helped mice maintain healthy conditions as they aged.

View Article: PubMed Central - PubMed

Affiliation: Research and Development, Benebios LLC, 10527 Garden Grove Blvd, Garden Grove, CA, 92843, USA. joon.lee@benebios.com.

ABSTRACT

Background: The gut microbiota is playing more important roles in host immune regulation than was initially expected. Since many benefits of microbes are highly strain-specific and their mechanistic details remain largely elusive, further identification of new probiotic bacteria with immunoregulatory potentials is of great interest.

Results: We have screened our collection of probiotic lactic acid bacteria (LAB) for their efficacy in modulating host immune response. Some LAB are characterized by suppression of TNF-α induction when LAB culture supernatants are added to THP-1 cells, demonstrating the LAB's anti-inflammatory potential. These suppressive materials were not inactivated by heat or trypsin. On the other hand, treatment of THP-1 directly with live bacterial cells identified a group of pro-inflammatory LAB, which stimulated significant production of TNF-α. Among those, we chose the Lactobacillus reuteri BM36301 as an anti-inflammatory strain and the L. reuteri BM36304 as a pro-inflammatory strain, and further studied their in vivo effects. We supplied C57BL/6 mice with these bacteria in drinking water while feeding them a standard diet for 20 weeks. Interestingly, these L. reuteri strains evoked different consequences depending on the gender of the mice. That is, males treated with anti-inflammatory BM36301 experienced less weight gain and higher testosterone level; females treated with BM36301 maintained lower serum TNF-α as well as healthy skin with active folliculogenesis and hair growth. Furthermore, while males treated with pro-inflammatory BM36304 developed higher serum levels of TNF-α and insulin, in contrast females did not experience such effects from this bacteria strain.

Conclusion: The L. reuteri BM36301 was selected as an anti-inflammatory strain in vitro. It helped mice maintain healthy conditions as they aged. These findings propose the L. reuteri BM36301 as a potential probiotic strain to improve various aspects of aging issues.

No MeSH data available.


Related in: MedlinePlus

Suppressive CMs are quantitative and resistant against heat and enzymatic digestion. a CM from L. reuteri strain BM36301 represses TNF-α production from LPS-treated THP-1 cells in a quantitative manner. TNF-α levels produced from the THP-1 cultures with (lanes 2 – 6) or without (lane 1) LPS were measured by ELISA method. For treatment of CM, 2.5 % (0.5×, lane 3), 5 % (1×, lane 4), 10 % (2×, lane 5), or 15 % (3×, lane 6) of THP-1 culture volume (v/v) was added at the time of LPS stimulation. CM from the control medium (MRS) was added to 5 % in lanes 1 and 2. The data show averages with SD from 3 independent assays. b CM from B. animalis subsp. lactis strain BM10307 contains metabolites that repress TNF-α production from LPS-treated THP-1 cells in a quantitative manner. The LPS and CM were treated as indicated. CM from the control medium (BL) was added to 5 % in lanes 1 and 2. Data are from 3 independent experiments. c CMs from BM36301 (gray) or BM10307 (black) were either boiled (lane 4) or treated with trypsin (lane 5) to compare their activities with native CMs (lane 3). Results are averages with SD from 3 independent experiments
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Fig2: Suppressive CMs are quantitative and resistant against heat and enzymatic digestion. a CM from L. reuteri strain BM36301 represses TNF-α production from LPS-treated THP-1 cells in a quantitative manner. TNF-α levels produced from the THP-1 cultures with (lanes 2 – 6) or without (lane 1) LPS were measured by ELISA method. For treatment of CM, 2.5 % (0.5×, lane 3), 5 % (1×, lane 4), 10 % (2×, lane 5), or 15 % (3×, lane 6) of THP-1 culture volume (v/v) was added at the time of LPS stimulation. CM from the control medium (MRS) was added to 5 % in lanes 1 and 2. The data show averages with SD from 3 independent assays. b CM from B. animalis subsp. lactis strain BM10307 contains metabolites that repress TNF-α production from LPS-treated THP-1 cells in a quantitative manner. The LPS and CM were treated as indicated. CM from the control medium (BL) was added to 5 % in lanes 1 and 2. Data are from 3 independent experiments. c CMs from BM36301 (gray) or BM10307 (black) were either boiled (lane 4) or treated with trypsin (lane 5) to compare their activities with native CMs (lane 3). Results are averages with SD from 3 independent experiments

Mentions: Since the CM from LAB is composed of various bacterial metabolites as well as cellular components, we sought to better understand the anti-inflammatory nature of the CM. The expression of TNF-α quantitatively diminished with increasing concentrations of the CMs from L. reuteri BM36301 (Fig. 2a) and B. animalis subsp. lactis BM10307 (Fig. 2b). Interestingly, a smaller amount of CM from BM36301 (0.5× input or 2.5 % v/v) actually induced TNF-α further, consistent with the observation that this CM contains TNF-α-inducing materials to some extent (Fig. 2a, lanes 2 and 3). Nonetheless, the suppressive effects became dominant in the face of higher amounts of CM, suggesting that the anti-inflammatory materials are quantitatively additive and that the corresponding receptors on the THP-1 cell surface are abundant enough to readily respond to increasing CM treatment. Overall, these observations suggest that the CMs contain active metabolites responsible for TNF-α suppression, in spite of their complex nature.Fig. 2


Characterization of the anti-inflammatory Lactobacillus reuteri BM36301 and its probiotic benefits on aged mice.

Lee J, Yang W, Hostetler A, Schultz N, Suckow MA, Stewart KL, Kim DD, Kim HS - BMC Microbiol. (2016)

Suppressive CMs are quantitative and resistant against heat and enzymatic digestion. a CM from L. reuteri strain BM36301 represses TNF-α production from LPS-treated THP-1 cells in a quantitative manner. TNF-α levels produced from the THP-1 cultures with (lanes 2 – 6) or without (lane 1) LPS were measured by ELISA method. For treatment of CM, 2.5 % (0.5×, lane 3), 5 % (1×, lane 4), 10 % (2×, lane 5), or 15 % (3×, lane 6) of THP-1 culture volume (v/v) was added at the time of LPS stimulation. CM from the control medium (MRS) was added to 5 % in lanes 1 and 2. The data show averages with SD from 3 independent assays. b CM from B. animalis subsp. lactis strain BM10307 contains metabolites that repress TNF-α production from LPS-treated THP-1 cells in a quantitative manner. The LPS and CM were treated as indicated. CM from the control medium (BL) was added to 5 % in lanes 1 and 2. Data are from 3 independent experiments. c CMs from BM36301 (gray) or BM10307 (black) were either boiled (lane 4) or treated with trypsin (lane 5) to compare their activities with native CMs (lane 3). Results are averages with SD from 3 independent experiments
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Fig2: Suppressive CMs are quantitative and resistant against heat and enzymatic digestion. a CM from L. reuteri strain BM36301 represses TNF-α production from LPS-treated THP-1 cells in a quantitative manner. TNF-α levels produced from the THP-1 cultures with (lanes 2 – 6) or without (lane 1) LPS were measured by ELISA method. For treatment of CM, 2.5 % (0.5×, lane 3), 5 % (1×, lane 4), 10 % (2×, lane 5), or 15 % (3×, lane 6) of THP-1 culture volume (v/v) was added at the time of LPS stimulation. CM from the control medium (MRS) was added to 5 % in lanes 1 and 2. The data show averages with SD from 3 independent assays. b CM from B. animalis subsp. lactis strain BM10307 contains metabolites that repress TNF-α production from LPS-treated THP-1 cells in a quantitative manner. The LPS and CM were treated as indicated. CM from the control medium (BL) was added to 5 % in lanes 1 and 2. Data are from 3 independent experiments. c CMs from BM36301 (gray) or BM10307 (black) were either boiled (lane 4) or treated with trypsin (lane 5) to compare their activities with native CMs (lane 3). Results are averages with SD from 3 independent experiments
Mentions: Since the CM from LAB is composed of various bacterial metabolites as well as cellular components, we sought to better understand the anti-inflammatory nature of the CM. The expression of TNF-α quantitatively diminished with increasing concentrations of the CMs from L. reuteri BM36301 (Fig. 2a) and B. animalis subsp. lactis BM10307 (Fig. 2b). Interestingly, a smaller amount of CM from BM36301 (0.5× input or 2.5 % v/v) actually induced TNF-α further, consistent with the observation that this CM contains TNF-α-inducing materials to some extent (Fig. 2a, lanes 2 and 3). Nonetheless, the suppressive effects became dominant in the face of higher amounts of CM, suggesting that the anti-inflammatory materials are quantitatively additive and that the corresponding receptors on the THP-1 cell surface are abundant enough to readily respond to increasing CM treatment. Overall, these observations suggest that the CMs contain active metabolites responsible for TNF-α suppression, in spite of their complex nature.Fig. 2

Bottom Line: On the other hand, treatment of THP-1 directly with live bacterial cells identified a group of pro-inflammatory LAB, which stimulated significant production of TNF-α.Furthermore, while males treated with pro-inflammatory BM36304 developed higher serum levels of TNF-α and insulin, in contrast females did not experience such effects from this bacteria strain.It helped mice maintain healthy conditions as they aged.

View Article: PubMed Central - PubMed

Affiliation: Research and Development, Benebios LLC, 10527 Garden Grove Blvd, Garden Grove, CA, 92843, USA. joon.lee@benebios.com.

ABSTRACT

Background: The gut microbiota is playing more important roles in host immune regulation than was initially expected. Since many benefits of microbes are highly strain-specific and their mechanistic details remain largely elusive, further identification of new probiotic bacteria with immunoregulatory potentials is of great interest.

Results: We have screened our collection of probiotic lactic acid bacteria (LAB) for their efficacy in modulating host immune response. Some LAB are characterized by suppression of TNF-α induction when LAB culture supernatants are added to THP-1 cells, demonstrating the LAB's anti-inflammatory potential. These suppressive materials were not inactivated by heat or trypsin. On the other hand, treatment of THP-1 directly with live bacterial cells identified a group of pro-inflammatory LAB, which stimulated significant production of TNF-α. Among those, we chose the Lactobacillus reuteri BM36301 as an anti-inflammatory strain and the L. reuteri BM36304 as a pro-inflammatory strain, and further studied their in vivo effects. We supplied C57BL/6 mice with these bacteria in drinking water while feeding them a standard diet for 20 weeks. Interestingly, these L. reuteri strains evoked different consequences depending on the gender of the mice. That is, males treated with anti-inflammatory BM36301 experienced less weight gain and higher testosterone level; females treated with BM36301 maintained lower serum TNF-α as well as healthy skin with active folliculogenesis and hair growth. Furthermore, while males treated with pro-inflammatory BM36304 developed higher serum levels of TNF-α and insulin, in contrast females did not experience such effects from this bacteria strain.

Conclusion: The L. reuteri BM36301 was selected as an anti-inflammatory strain in vitro. It helped mice maintain healthy conditions as they aged. These findings propose the L. reuteri BM36301 as a potential probiotic strain to improve various aspects of aging issues.

No MeSH data available.


Related in: MedlinePlus