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Gut microbiome derived metabolites modulate intestinal epithelial cell damage and mitigate Graft-versus-Host Disease

View Article: PubMed Central - PubMed

ABSTRACT

The impact of alterations in intestinal microbiota on microbial metabolites and on disease processes, such as graft-versus-host disease (GVHD), is not known. Here we performed unbiased analysis to identify novel alterations in gastrointestinal microbiota-derived short chain fatty acids (SCFA) after allogeneic bone marrow transplant (allo-BMT). Alterations in the amounts of only one SCFA, butyrate, were observed only within the intestinal tissue. The reduced butyrate in CD326+ intestinal epithelial cells (IECs) after allo-BMT resulted in decreased histone acetylation, which was restored upon local administration of exogenous butyrate. Butyrate restoration improved IEC junctional integrity, decreased apoptosis, and mitigated GVHD. Furthermore, alteration of the indigenous microbiota with 17 rationally selected strains of high butyrate producing Clostridia also decreased GVHD. These data demonstrate a heretofore unrecognized role of microbial metabolites and suggest that local and specific alteration of microbial metabolites has direct salutary effects on GVHD target tissues and can mitigate its severity.

No MeSH data available.


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Donor Treg cells are dispensable for GVHD protection(a)-(c) Intestinal immunophenotypical analysis of recipients 21 days following allogeneic (C57BL/6J → BALB/c) BMT treated with either butyrate or vehicle via intragastric gavage. (a) Total cell numbers of intestinal CD4+ & CD8+ T cells (left column) and activated T cells: CD69+ T cells (middle column) & CD44hi T cells (right column). (b) Ratio of intestinal Treg cells (CD4+CD25+FoxP3+) to effector cells (CD4+FoxP3−) and (c) total cell number of intestinal donor macrophages (CD11b+F4.80+) in recipients of allo-BMT treated with vehicle and butyrate. (d)-(e) Recipients received CD4+CD25− (Treg depleted) 1.5 × 106 donor T cells and T cell depleted (TCD) bone marrow. (d) GVHD score and (e) survival in animals treated with vehicle or butyrate (10mg/kg); syngeneic n=6, n=12 mice per allogeneic group. (f)-(g) Recipients received Treg depleted (C57BL/6J DT.Treg) 0.5 × 106 donor cells and TCD bone marrow with resulting (f) GVHD score and (g) survival in butyrate treated BMT recipients. (h) HDAC activity in T cells of the spleen 21 days following syngeneic or allogeneic BMT. Syngeneic n=5, n = 10 mice per allogeneic group. *P < .05; **P < .01; ***P < .0001 of students t-test a - c; Mantel-Cox log-rank test d - g. Bars and error bars represent the means and standard errors of the mean, respectively.
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Figure 4: Donor Treg cells are dispensable for GVHD protection(a)-(c) Intestinal immunophenotypical analysis of recipients 21 days following allogeneic (C57BL/6J → BALB/c) BMT treated with either butyrate or vehicle via intragastric gavage. (a) Total cell numbers of intestinal CD4+ & CD8+ T cells (left column) and activated T cells: CD69+ T cells (middle column) & CD44hi T cells (right column). (b) Ratio of intestinal Treg cells (CD4+CD25+FoxP3+) to effector cells (CD4+FoxP3−) and (c) total cell number of intestinal donor macrophages (CD11b+F4.80+) in recipients of allo-BMT treated with vehicle and butyrate. (d)-(e) Recipients received CD4+CD25− (Treg depleted) 1.5 × 106 donor T cells and T cell depleted (TCD) bone marrow. (d) GVHD score and (e) survival in animals treated with vehicle or butyrate (10mg/kg); syngeneic n=6, n=12 mice per allogeneic group. (f)-(g) Recipients received Treg depleted (C57BL/6J DT.Treg) 0.5 × 106 donor cells and TCD bone marrow with resulting (f) GVHD score and (g) survival in butyrate treated BMT recipients. (h) HDAC activity in T cells of the spleen 21 days following syngeneic or allogeneic BMT. Syngeneic n=5, n = 10 mice per allogeneic group. *P < .05; **P < .01; ***P < .0001 of students t-test a - c; Mantel-Cox log-rank test d - g. Bars and error bars represent the means and standard errors of the mean, respectively.

Mentions: Treg cells mitigate GVHD and butyrate has been shown to increase intestinal Treg cells17,28. We therefore analyzed the cellular contents of the intestine 21 days after allo-BMT to determine whether butyrate had an impact on local Treg cells. The total numbers of CD45.1+ cells recovered from the intestinal lamina propria were not different between vehicle- and butyrate-treated allo-BMT recipients (Supplementary Fig. 4g). By contrast, intestinal infiltration of donor CD4+ and CD8+ T cells and activated T cells (CD69+ or CD44hi) was decreased in animals that received local intragastric butyrate administration (Fig. 4a). However, the ratio of donor Treg cells to effector T cells was not different in the intestines of these animals (Fig. 4b). Microbiota-derived butyrate has been shown to increase immune-regulatory macrophages in the GI tract which increased Treg cells29. However we observed no difference in the total number of donor macrophages in the intestine of allogenenic recipients that were treated with either vehicle or butyrate (Fig. 4c).


Gut microbiome derived metabolites modulate intestinal epithelial cell damage and mitigate Graft-versus-Host Disease
Donor Treg cells are dispensable for GVHD protection(a)-(c) Intestinal immunophenotypical analysis of recipients 21 days following allogeneic (C57BL/6J → BALB/c) BMT treated with either butyrate or vehicle via intragastric gavage. (a) Total cell numbers of intestinal CD4+ & CD8+ T cells (left column) and activated T cells: CD69+ T cells (middle column) & CD44hi T cells (right column). (b) Ratio of intestinal Treg cells (CD4+CD25+FoxP3+) to effector cells (CD4+FoxP3−) and (c) total cell number of intestinal donor macrophages (CD11b+F4.80+) in recipients of allo-BMT treated with vehicle and butyrate. (d)-(e) Recipients received CD4+CD25− (Treg depleted) 1.5 × 106 donor T cells and T cell depleted (TCD) bone marrow. (d) GVHD score and (e) survival in animals treated with vehicle or butyrate (10mg/kg); syngeneic n=6, n=12 mice per allogeneic group. (f)-(g) Recipients received Treg depleted (C57BL/6J DT.Treg) 0.5 × 106 donor cells and TCD bone marrow with resulting (f) GVHD score and (g) survival in butyrate treated BMT recipients. (h) HDAC activity in T cells of the spleen 21 days following syngeneic or allogeneic BMT. Syngeneic n=5, n = 10 mice per allogeneic group. *P < .05; **P < .01; ***P < .0001 of students t-test a - c; Mantel-Cox log-rank test d - g. Bars and error bars represent the means and standard errors of the mean, respectively.
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Related In: Results  -  Collection

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Figure 4: Donor Treg cells are dispensable for GVHD protection(a)-(c) Intestinal immunophenotypical analysis of recipients 21 days following allogeneic (C57BL/6J → BALB/c) BMT treated with either butyrate or vehicle via intragastric gavage. (a) Total cell numbers of intestinal CD4+ & CD8+ T cells (left column) and activated T cells: CD69+ T cells (middle column) & CD44hi T cells (right column). (b) Ratio of intestinal Treg cells (CD4+CD25+FoxP3+) to effector cells (CD4+FoxP3−) and (c) total cell number of intestinal donor macrophages (CD11b+F4.80+) in recipients of allo-BMT treated with vehicle and butyrate. (d)-(e) Recipients received CD4+CD25− (Treg depleted) 1.5 × 106 donor T cells and T cell depleted (TCD) bone marrow. (d) GVHD score and (e) survival in animals treated with vehicle or butyrate (10mg/kg); syngeneic n=6, n=12 mice per allogeneic group. (f)-(g) Recipients received Treg depleted (C57BL/6J DT.Treg) 0.5 × 106 donor cells and TCD bone marrow with resulting (f) GVHD score and (g) survival in butyrate treated BMT recipients. (h) HDAC activity in T cells of the spleen 21 days following syngeneic or allogeneic BMT. Syngeneic n=5, n = 10 mice per allogeneic group. *P < .05; **P < .01; ***P < .0001 of students t-test a - c; Mantel-Cox log-rank test d - g. Bars and error bars represent the means and standard errors of the mean, respectively.
Mentions: Treg cells mitigate GVHD and butyrate has been shown to increase intestinal Treg cells17,28. We therefore analyzed the cellular contents of the intestine 21 days after allo-BMT to determine whether butyrate had an impact on local Treg cells. The total numbers of CD45.1+ cells recovered from the intestinal lamina propria were not different between vehicle- and butyrate-treated allo-BMT recipients (Supplementary Fig. 4g). By contrast, intestinal infiltration of donor CD4+ and CD8+ T cells and activated T cells (CD69+ or CD44hi) was decreased in animals that received local intragastric butyrate administration (Fig. 4a). However, the ratio of donor Treg cells to effector T cells was not different in the intestines of these animals (Fig. 4b). Microbiota-derived butyrate has been shown to increase immune-regulatory macrophages in the GI tract which increased Treg cells29. However we observed no difference in the total number of donor macrophages in the intestine of allogenenic recipients that were treated with either vehicle or butyrate (Fig. 4c).

View Article: PubMed Central - PubMed

ABSTRACT

The impact of alterations in intestinal microbiota on microbial metabolites and on disease processes, such as graft-versus-host disease (GVHD), is not known. Here we performed unbiased analysis to identify novel alterations in gastrointestinal microbiota-derived short chain fatty acids (SCFA) after allogeneic bone marrow transplant (allo-BMT). Alterations in the amounts of only one SCFA, butyrate, were observed only within the intestinal tissue. The reduced butyrate in CD326+ intestinal epithelial cells (IECs) after allo-BMT resulted in decreased histone acetylation, which was restored upon local administration of exogenous butyrate. Butyrate restoration improved IEC junctional integrity, decreased apoptosis, and mitigated GVHD. Furthermore, alteration of the indigenous microbiota with 17 rationally selected strains of high butyrate producing Clostridia also decreased GVHD. These data demonstrate a heretofore unrecognized role of microbial metabolites and suggest that local and specific alteration of microbial metabolites has direct salutary effects on GVHD target tissues and can mitigate its severity.

No MeSH data available.


Related in: MedlinePlus