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. 2012;8(5):e1002714.
doi: 10.1371/journal.ppat.1002714. Epub 2012 May 31.

Probiotic Bifidobacterium breve induces IL-10-producing Tr1 cells in the colon

Seong Gyu Jeon  1 , Hisako Kayama, Yoshiyasu Ueda, Takuya Takahashi, Takashi Asahara, Hirokazu Tsuji, Noriko M Tsuji, Hiroshi Kiyono, Ji Su Ma, Takashi Kusu, Ryu Okumura, Hiromitsu Hara, Hiroki Yoshida, Masahiro Yamamoto, Koji Nomoto, Kiyoshi Takeda
Affiliations

Probiotic Bifidobacterium breve induces IL-10-producing Tr1 cells in the colon

Seong Gyu Jeon et al. PLoS Pathog. 2012.

Abstract

Specific intestinal microbiota has been shown to induce Foxp3(+) regulatory T cell development. However, it remains unclear how development of another regulatory T cell subset, Tr1 cells, is regulated in the intestine. Here, we analyzed the role of two probiotic strains of intestinal bacteria, Lactobacillus casei and Bifidobacterium breve in T cell development in the intestine. B. breve, but not L. casei, induced development of IL-10-producing Tr1 cells that express cMaf, IL-21, and Ahr in the large intestine. Intestinal CD103(+) dendritic cells (DCs) mediated B. breve-induced development of IL-10-producing T cells. CD103(+) DCs from Il10(-/-), Tlr2(-/-), and Myd88(-/-) mice showed defective B. breve-induced Tr1 cell development. B. breve-treated CD103(+) DCs failed to induce IL-10 production from co-cultured Il27ra(-/-) T cells. B. breve treatment of Tlr2(-/-) mice did not increase IL-10-producing T cells in the colonic lamina propria. Thus, B VSports手机版. breve activates intestinal CD103(+) DCs to produce IL-10 and IL-27 via the TLR2/MyD88 pathway thereby inducing IL-10-producing Tr1 cells in the large intestine. Oral B. breve administration ameliorated colitis in immunocompromised mice given naïve CD4(+) T cells from wild-type mice, but not Il10(-/-) mice. These findings demonstrate that B. breve prevents intestinal inflammation through the induction of intestinal IL-10-producing Tr1 cells. .

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Induction of IL-10-producing CD4+ T cells by B. breve in the colonic lamina propria.
6-week-old C57BL/6 mice were fed L. casei or B. breve or placebo daily (each, 1×109) by oral gavage for 3 months (n = 8). Intestinal lamina propria lymphocytes were analyzed for cytokine production by flow cytometry. Percentages of IL-10-, IL-17-, and IFN-γ-producing CD4+ T cells of mice administered with L. casei (A) or B. breve (C) were shown. *P<0.05. (B, D) Representative FACS dot plots showing production of IL-10 and IFN-γ gated on colonic CD4+ T cells in the indicated mice.
Figure 2
Figure 2. Induction of Foxp3− IL-10-producing CD4+ T cells by B. breve.
6-week-old BALB/c mice were administered with B. breve or placebo orally for 1–4 weeks (n = 5). At the indicated time point, mice were sacrificed and CD4+ T cells in the colonic lamina propria were analyzed by flow cytometry. Percentages of CD4+ IL-10+ T cells (A), and CD4+ Foxp3+ T cells (B) are shown. *P<0.02. Representative FACS dot plots for IL-10+ and IL-17+ T cells (C), and histogram for Foxp3+ T cells (D) gated on CD4+ T cells are shown. (E) Foxp3 GFP mice were fed with B. breve for 4 weeks. CD4+ T cells in the colonic lamina propria were analyzed for expression of GFP and IL-10. (F) Percentages of IL-10+ T cells in Foxp3+ or Foxp3− CD4+ T cells (n = 5). Data are representative of two independent experiments; means ± S.D. *P<0.02.
Figure 3
Figure 3. Intestinal DCs mediate B. breve-dependent Tr1 cell development.
CD11c+ DCs (5×104) were isolated from the colonic lamina propria, and cultured with B. breve, L. casei, B. adolescentis, B. longum, or B. bifidum (5×104) for 24 h. After washing, DCs were co-cultured with splenic naïve CD4+ T cells (5×104) in the presence of soluble anti-CD3 mAb for 4 days. (A) T cells were harvested and re-stimulated with plate-bound anti-CD3 and soluble anti-CD28 mAbs for 24 h. IL-10 concentrations in the culture supernatants were analyzed by ELISA. *P<0.001. (B) T cells were harvested and re-stimulated with plate-bound anti-CD3 and soluble anti-CD28 mAbs for 24 h. IL-10 concentrations in the culture supernatants were analyzed by ELISA. *P<0.001. (C) T cells were collected, and then stained for CD4 and Foxp3. Foxp3 expression in CD4+ cells is shown. (D) T cells were harvested, and stimulated with anti-CD3 and anti-CD28 mAbs for 4 h. Total RNA was then extracted to analyze expression of cMaf, Il21, and Ahr by quantitative real-time RT-PCR. Data are representative of five independent experiments and show mean values ± S.D. of triplicate determinations. *P<0.05, **P<0.01.
Figure 4
Figure 4. CD103+ DCs, but not CX3CR1+ DCs, induce B. breve-dependent Tr1 cell differentiation.
CD103+ CX3CR1− CD11b− CD11c+ DCs (CD103+ DCs) and CX3CR1+ CD11b+ CD11c+ DCs (CX3CR1+ DCs) were isolated from the colonic lamina propria, and treated with the same numbers of B. breve for 24 h. After washing, splenic naïve CD4+ T cells were co-cultured with B. breve-treated CD103+ DCs or CX3CR1+ DCs in the presence of anti-CD3 mAb for 4 days. (A) T cells were then harvested and re-stimulated for 24 h to analyze IL-10 production by ELISA. *P<0.05. (B) T cells were collected, and re-stimulated with PMA and ionophore for 8 h. Intracellular expression of Foxp3 and IL-10 was then analyzed by flow cytometry. (C) C57BL/6J mice (n = 5) were fed with B. breve for 3 weeks. Then, CD103+ DCs were isolated from MLN and the colonic lamina propria, and co-cultured with splenic naïve CD4+ T cells. The co-cultured T cells were re-stimulated and IL-10 concentration in the supernatants was analyzed by ELISA. Data are representative of four independent experiments and show mean values ± S.D. of triplicate determinations. *P<0.05, **P<0.01.
Figure 5
Figure 5. IL-10/IL-27-dependent Tr1 cell development by B. breve-treated DCs.
(A) CD103+ DCs were isolated from the colonic lamina propria, and incubated with B. breve for 4 h. Total RNA was extracted and analyzed for mRNA expression of Il27p28, Ebi3, and Il10 by quantitative real-time RT-PCR. N.D, not detected. **P<0.01, ***P<0.001. (B) Naïve T cells were co-cultured with B. breve-treated CD103+ DC in the presence of the indicated neutralizing antibody for 4 days. T cells were then harvested and re-stimulated with anti-CD3 and CD28 mAbs for 24 h. IL-10 concentrations in the supernatants were measured by ELISA. *P<0.05, **P<0.01, ***P<0.001, N.S, not significant. (C) CD103+ DCs were isolated from the colonic lamina propria of wild-type and Il10 −/− mice (C57BL/6 background) and incubated with B. breve. Naïve CD4+ T cells from wild-type C57BL/6 mice were then co-cultured with B. breve-treated DCs. T cell production of IL-10 was analyzed by ELISA. N.D, not detected. (D) CD103+ DCs were isolated from the colonic lamina propria of wild-type BALB/c mice and incubated with B. breve. Naïve CD4+ T cells from wild-type and Il27ra −/− mice (BALB/c background) were then co-cultured with B. breve-treated DCs. T cell production of IL-10 was measured by ELISA. Data are representative of three independent experiments and show mean values ± S.D. of triplicate determinations. *P<0.05.
Figure 6
Figure 6. B. breve induces Tr1 cells in a TLR2/MyD88-dependent manner.
(A, C) CD103+ DCs were isolated from the colonic lamina propria of wild-type, Myd88 −/− (A) and Tlr2 −/− (C) mice, incubated with B. breve for 4 h, and then analyzed for mRNA expression of Il27p28, Ebi3, and Il10. *P<0.05, **P<0.01. (B, D) Wild-type, Myd88 −/− (B) and Tlr2 −/− (D) CD103+ DCs were incubated with B. breve for 24 h, and then co-cultured with naïve CD4+ T cells from wild-type mice for 4 days. T cells were harvested and re-stimulated for 24 h. IL-10 production in the supernatants was analyzed by ELISA. Data are representative of three independent experiments and show mean values ± S.D. of triplicate determinations. *P<0.05. N.D, not detected. (E) 6-week-old wild-type and Tlr2 −/− mice (BALB/c background) were fed with B. breve or placebo for 3 weeks (n = 5). Then, the mice were sacrificed and colonic lamina propria lymphocytes were analyzed for IL-10 production by flow cytometry. The percentage of IL-10+ cells gated on CD4+ T cells is shown in the indicated mice. Data are representative of three independent experiments and show mean values ± S.D. of triplicate determinations. *P<0.05. (F) Representative FACS plots of IL-10- and IFN-γ-producing CD4+ T cells were shown.
Figure 7
Figure 7. IL-10-dependent amelioration of intestinal inflammation by B. breve.
(A, C) 6 week-old SCID mice (n = 8 per group) were intraperitoneally injected with PBS or 3×105 of naïve CD4+ T cells from wild-type BALB/c mice (A) or Il10 −/− mice (BALB/c background) (B). The mice were orally administered daily with B. breve from 1 week before the T cell transfer to the end of experiment. Changes in body weight were monitored daily and presented relative to initial body weight. *P<0.05, Error bars, S.E.M. (C) Production of IL-10, IL-17 and IFN-γ from the colon of wild-type T cell-transferred SCID mice daily administered with B. breve or placebo (n = 5 per group). *P<0.0064, **P<0.0005. (D) Hematoxylin and eosin staining of colon sections at 4 weeks after the transfer. Original magnification, ×400. (E) Clinical scores for colitis were shown in the indicated group. Data are representative of two independent experiments. *P<0.05, **P<0.01. N.S, not significant.
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References

    1. Frank DN, St Amand AL, Feldman RA, Boedeker EC, Harpaz N, et al. Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases. Proc Natl Acad Sci U S A. 2007;104:13780–13785. - PMC (V体育2025版) - PubMed
    1. Peterson DA, Frank DN, Pace NR, Gordon JI. Metagenomic approaches for defining the pathogenesis of inflammatory bowel diseases. Cell Host Microbe. 2008;3:417–427. - PMC - PubMed
    1. Qin J, Li R, Raes J, Arumugam M, Burgdorf KS, et al. A human gut microbial gene catalogue established by metagenomic sequencing. Nature. 2010;464:59–65. - VSports手机版 - PMC - PubMed
    1. Elinav E, Strowig T, Kau AL, Henao-Mejia J, Thaiss CA, et al. NLRP6 inflammasome regulates colonic microbial ecology and risk for colitis. Cell. 2011;145:745–757. - PMC - PubMed
    1. Hart AL, Stagg AJ, Kamm MA. Use of probiotics in the treatment of inflammatory bowel disease. J Clin Gastroenterol. 2003;36:111–119. - PubMed

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