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Extracellular matrix-associated cytokines regulate CD4+ effector T-cell responses in the human intestinal mucosa.

Huff KR, Akhtar LN, Fox AL, Cannon JA, Smith PD, Smythies LE - Mucosal Immunol (2011)

Bottom Line: In this study, we uncovered a role for intestinal stromal products in the innate regulation of effector T cells.Stroma-conditioned media (S-CM) derived from the normal human intestinal stroma (transforming growth factor-β (TGF-β)(hi)/interleukin (IL)-6(lo)/IL-1β(lo)) significantly downregulated T-cell proliferation and interferon-γ (IFN-γ) production compared with S-CM derived from the inflamed Crohn's mucosa (TGF-β(hi)/IL-6(hi)/IL-1β(hi)).These findings implicate stromal TGF-β in the downregulation of T-cell 2 responses in the normal intestinal mucosa, and stromal IL-6 and IL-1β in the promotion of Th1 and Th17 responses in the inflamed Crohn's mucosa, suggesting an innate regulatory function for the intestinal extracellular matrix.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine (Gastroenterology), University of Alabama at Birmingham, Birmingham, Alabama, USA.

ABSTRACT
Extracellular matrix (stroma) regulation of mucosal T-cell function is incompletely understood. In this study, we uncovered a role for intestinal stromal products in the innate regulation of effector T cells. Stroma-conditioned media (S-CM) derived from the normal human intestinal stroma (transforming growth factor-β (TGF-β)(hi)/interleukin (IL)-6(lo)/IL-1β(lo)) significantly downregulated T-cell proliferation and interferon-γ (IFN-γ) production compared with S-CM derived from the inflamed Crohn's mucosa (TGF-β(hi)/IL-6(hi)/IL-1β(hi)). Antibody neutralization studies showed that TGF-β in normal S-CM inhibited T-cell proliferation and IFN-γ production, whereas IL-6 plus IL-1β in Crohn's S-CM promoted T-cell proliferation, and IL-1β alone promoted IFN-γ and IL-17 release. Importantly, normal S-CM inhibited T-bet expression, whereas Crohn's S-CM activated signal transducer and activator of transcription 3, suggesting that discordant T-cell responses are regulated at the transcription factor and signaling levels. These findings implicate stromal TGF-β in the downregulation of T-cell 2 responses in the normal intestinal mucosa, and stromal IL-6 and IL-1β in the promotion of Th1 and Th17 responses in the inflamed Crohn's mucosa, suggesting an innate regulatory function for the intestinal extracellular matrix.

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Crohn’s S-CM promotes IL-17 release by CD4 T-cells in an IL-1β -dependent manner(A) IL-17 release by CD3/CD28-stimulated T-cells cultured 4 days in the presence of normal S-CM or Crohn’s S-CM compared to stimulated T-cells cultured in media alone (n=3). Inset shows results from a representative experiment (n=3) with IL-17 release (pg/mL) by CD3/CD28-stimulated blood T-cells cultured for 4 days in the presence of normal or Crohn’s S-CM. (B) IL-17 release by CD3/CD28-stimulated T-cells cultured for 4 days in Crohn’s S-CM ± anti-IL-6 antibody and/or anti-IL-1β antibodies (1 µg/mL) (n=2, each performed in triplicate). Error bars indicate the standard error of the mean. *p ≤ 0.05 and ** p ≤ 0.01.
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Figure 5: Crohn’s S-CM promotes IL-17 release by CD4 T-cells in an IL-1β -dependent manner(A) IL-17 release by CD3/CD28-stimulated T-cells cultured 4 days in the presence of normal S-CM or Crohn’s S-CM compared to stimulated T-cells cultured in media alone (n=3). Inset shows results from a representative experiment (n=3) with IL-17 release (pg/mL) by CD3/CD28-stimulated blood T-cells cultured for 4 days in the presence of normal or Crohn’s S-CM. (B) IL-17 release by CD3/CD28-stimulated T-cells cultured for 4 days in Crohn’s S-CM ± anti-IL-6 antibody and/or anti-IL-1β antibodies (1 µg/mL) (n=2, each performed in triplicate). Error bars indicate the standard error of the mean. *p ≤ 0.05 and ** p ≤ 0.01.

Mentions: STAT3 activation plays a critical role in the development of IL-17-producing T-cells 16. Therefore, we determined whether the discordant regulation of T-cell function by normal versus Crohn’s stromal factors extended to IL-17 secretion. In the presence of normal S-CM, CD3/CD28-stimulated T-cell production of IL-17 was reduced, albeit not significantly, compared to control T-cells (Figure 5A). However, in the presence of increasing concentrations of Crohn’s S-CM, T-cells secreted markedly higher levels of IL-17 compared to T-cells cultured in media alone (p < 0.01), with similar amounts of IL-17 released at all concentrations of Crohn’s S-CM (Figure 5A and inset). Since the proportion of pro-inflammatory (IL-6 and IL-1β) to down-regulatory (TGF-β) cytokines was constant at each concentration of Crohn’s S-CM, these findings suggest that the relative proportion of the cytokines was more important than their absolute concentration in the regulation of IL-17 secretion in the presence of Crohn’s S-CM.


Extracellular matrix-associated cytokines regulate CD4+ effector T-cell responses in the human intestinal mucosa.

Huff KR, Akhtar LN, Fox AL, Cannon JA, Smith PD, Smythies LE - Mucosal Immunol (2011)

Crohn’s S-CM promotes IL-17 release by CD4 T-cells in an IL-1β -dependent manner(A) IL-17 release by CD3/CD28-stimulated T-cells cultured 4 days in the presence of normal S-CM or Crohn’s S-CM compared to stimulated T-cells cultured in media alone (n=3). Inset shows results from a representative experiment (n=3) with IL-17 release (pg/mL) by CD3/CD28-stimulated blood T-cells cultured for 4 days in the presence of normal or Crohn’s S-CM. (B) IL-17 release by CD3/CD28-stimulated T-cells cultured for 4 days in Crohn’s S-CM ± anti-IL-6 antibody and/or anti-IL-1β antibodies (1 µg/mL) (n=2, each performed in triplicate). Error bars indicate the standard error of the mean. *p ≤ 0.05 and ** p ≤ 0.01.
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Related In: Results  -  Collection

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Figure 5: Crohn’s S-CM promotes IL-17 release by CD4 T-cells in an IL-1β -dependent manner(A) IL-17 release by CD3/CD28-stimulated T-cells cultured 4 days in the presence of normal S-CM or Crohn’s S-CM compared to stimulated T-cells cultured in media alone (n=3). Inset shows results from a representative experiment (n=3) with IL-17 release (pg/mL) by CD3/CD28-stimulated blood T-cells cultured for 4 days in the presence of normal or Crohn’s S-CM. (B) IL-17 release by CD3/CD28-stimulated T-cells cultured for 4 days in Crohn’s S-CM ± anti-IL-6 antibody and/or anti-IL-1β antibodies (1 µg/mL) (n=2, each performed in triplicate). Error bars indicate the standard error of the mean. *p ≤ 0.05 and ** p ≤ 0.01.
Mentions: STAT3 activation plays a critical role in the development of IL-17-producing T-cells 16. Therefore, we determined whether the discordant regulation of T-cell function by normal versus Crohn’s stromal factors extended to IL-17 secretion. In the presence of normal S-CM, CD3/CD28-stimulated T-cell production of IL-17 was reduced, albeit not significantly, compared to control T-cells (Figure 5A). However, in the presence of increasing concentrations of Crohn’s S-CM, T-cells secreted markedly higher levels of IL-17 compared to T-cells cultured in media alone (p < 0.01), with similar amounts of IL-17 released at all concentrations of Crohn’s S-CM (Figure 5A and inset). Since the proportion of pro-inflammatory (IL-6 and IL-1β) to down-regulatory (TGF-β) cytokines was constant at each concentration of Crohn’s S-CM, these findings suggest that the relative proportion of the cytokines was more important than their absolute concentration in the regulation of IL-17 secretion in the presence of Crohn’s S-CM.

Bottom Line: In this study, we uncovered a role for intestinal stromal products in the innate regulation of effector T cells.Stroma-conditioned media (S-CM) derived from the normal human intestinal stroma (transforming growth factor-β (TGF-β)(hi)/interleukin (IL)-6(lo)/IL-1β(lo)) significantly downregulated T-cell proliferation and interferon-γ (IFN-γ) production compared with S-CM derived from the inflamed Crohn's mucosa (TGF-β(hi)/IL-6(hi)/IL-1β(hi)).These findings implicate stromal TGF-β in the downregulation of T-cell 2 responses in the normal intestinal mucosa, and stromal IL-6 and IL-1β in the promotion of Th1 and Th17 responses in the inflamed Crohn's mucosa, suggesting an innate regulatory function for the intestinal extracellular matrix.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine (Gastroenterology), University of Alabama at Birmingham, Birmingham, Alabama, USA.

ABSTRACT
Extracellular matrix (stroma) regulation of mucosal T-cell function is incompletely understood. In this study, we uncovered a role for intestinal stromal products in the innate regulation of effector T cells. Stroma-conditioned media (S-CM) derived from the normal human intestinal stroma (transforming growth factor-β (TGF-β)(hi)/interleukin (IL)-6(lo)/IL-1β(lo)) significantly downregulated T-cell proliferation and interferon-γ (IFN-γ) production compared with S-CM derived from the inflamed Crohn's mucosa (TGF-β(hi)/IL-6(hi)/IL-1β(hi)). Antibody neutralization studies showed that TGF-β in normal S-CM inhibited T-cell proliferation and IFN-γ production, whereas IL-6 plus IL-1β in Crohn's S-CM promoted T-cell proliferation, and IL-1β alone promoted IFN-γ and IL-17 release. Importantly, normal S-CM inhibited T-bet expression, whereas Crohn's S-CM activated signal transducer and activator of transcription 3, suggesting that discordant T-cell responses are regulated at the transcription factor and signaling levels. These findings implicate stromal TGF-β in the downregulation of T-cell 2 responses in the normal intestinal mucosa, and stromal IL-6 and IL-1β in the promotion of Th1 and Th17 responses in the inflamed Crohn's mucosa, suggesting an innate regulatory function for the intestinal extracellular matrix.

Show MeSH
Related in: MedlinePlus