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Transforming growth factor (TGF)-beta1-producing regulatory T cells induce Smad-mediated interleukin 10 secretion that facilitates coordinated immunoregulatory activity and amelioration of TGF-beta1-mediated fibrosis.

Kitani A, Fuss I, Nakamura K, Kumaki F, Usui T, Strober W - J. Exp. Med. (2003)

Bottom Line: In addition, using a self-inactivating retrovirus luciferase reporter construct we showed that TGF-beta1 induces Smad4, which then binds to and activates the IL-10 promoter.Furthermore, intranasal TGF-beta1 plasmid administration ameliorates bleomycin-induced fibrosis in wild-type but not IL-10-deficient mice, strongly suggesting that the amelioration is IL-10 dependent and that IL-10 protects mice from TGF-beta1-mediated fibrosis.Taken together, these findings suggest that the induction of IL-10 by TGF-beta1 is not fortuitous, but instead fulfills important requirements of TGF-beta1 function after its secretion by regulatory T cells.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 10, Room 11N238, 10 Center Drive, Bethesda, MD 20892, USA.

ABSTRACT
Interleukin (IL)-10 and transforming growth factor (TGF)-beta1 are suppressor cytokines that frequently occur together during a regulatory T cell response. Here we used a one gene doxycycline (Dox)-inducible plasmid encoding TGF-beta1 to analyze this association and test its utility. In initial studies, we showed that intranasal administration of this plasmid (along with Dox) led to the appearance of TGF-beta1-producing cells (in spleen and lamina propria) and the almost concomitant appearance of IL-10-producing cells. Moreover, we showed that these cells exert Dox-regulated suppression of the T helper cell (Th)1-mediated inflammation in trinitrobenzene sulfonic acid colitis. In subsequent in vitro studies using retroviral TGF-beta1 expression, we established that IL-10 production by Th1 cells occurs after exposure to TGF-beta1 from either an endogenous or exogenous source. In addition, using a self-inactivating retrovirus luciferase reporter construct we showed that TGF-beta1 induces Smad4, which then binds to and activates the IL-10 promoter. Furthermore, intranasal TGF-beta1 plasmid administration ameliorates bleomycin-induced fibrosis in wild-type but not IL-10-deficient mice, strongly suggesting that the amelioration is IL-10 dependent and that IL-10 protects mice from TGF-beta1-mediated fibrosis. Taken together, these findings suggest that the induction of IL-10 by TGF-beta1 is not fortuitous, but instead fulfills important requirements of TGF-beta1 function after its secretion by regulatory T cells.

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Effect of intranasal pTet-On-TGF-β1 administration on TNBS-induced colitis. (a) Weight changes of mice groups administered intrarectal TNBS and intranasal pTet-On-TGF-β or pTet-On-mock (control plasmid) with or without Dox (i.p.) Each group, n = 10. (b) LP cells from mice 5 d after TNBS colitis induction were stimulated and cultured for ELISA assays of TGF-β1, IFN-γ, IL-10, and IL-12. As shown, LP cells from mice administered Dox plus pTet-On-TGF-β1 produced large amounts of TGF-β1 and IL-10 and exhibited suppressed IL-12 and IFN-γ production. Data shown is representative of three independent experiments.
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fig2: Effect of intranasal pTet-On-TGF-β1 administration on TNBS-induced colitis. (a) Weight changes of mice groups administered intrarectal TNBS and intranasal pTet-On-TGF-β or pTet-On-mock (control plasmid) with or without Dox (i.p.) Each group, n = 10. (b) LP cells from mice 5 d after TNBS colitis induction were stimulated and cultured for ELISA assays of TGF-β1, IFN-γ, IL-10, and IL-12. As shown, LP cells from mice administered Dox plus pTet-On-TGF-β1 produced large amounts of TGF-β1 and IL-10 and exhibited suppressed IL-12 and IFN-γ production. Data shown is representative of three independent experiments.

Mentions: In a further study we investigated the ability of the Tet-On-TGF-β1 plasmid to affect an inflammatory state, exemplified by the Th1-type colitis induced by TNBS (4–7). Four groups of mice were studied: those receiving pTet-On-TGF-β1 with or without Dox and those receiving control plasmid (pTet-On-mock) also with or without Dox. As shown in Fig. 2 a, using body weight as an indicator of colitis, it was clear that the group that received pTet-On-TGF-β1 (i.n.) and Dox (i.p.) was protected from developing colitis, whereas the group receiving the plasmid in the absence of Dox (or those mice receiving pTet-ON-mock again with and without Dox) developed colitis. In addition, although LP cells from all three groups developing colitis produced large amounts of IL-12 and IFN-γ and small amounts of TGF-β1, LP cells from mice prevented from developing colitis produced small amounts of IL-12 and IFN-γ and very large amounts of TGF-β1 (Fig. 2 b). Of interest, the latter was much greater than that produced by cells taken from mice without inflammation, reflecting the activation of the CMV promoter in an inflammatory milieu. In addition, LP cells from this group, but not those from the groups in which the plasmid was not accompanied by Dox administration, produced large amounts of IL-10. Thus, it was obvious that such IL-10 secretion was due to plasmid-induced TGF-β1 and not simply to the presence of a Th1 inflammation. In further studies not shown, we found that intranasal administration of pTet-On-TGF-β1 also reversed TNBS colitis after inflammation was established, but only in the presence of concomitant Dox administration.


Transforming growth factor (TGF)-beta1-producing regulatory T cells induce Smad-mediated interleukin 10 secretion that facilitates coordinated immunoregulatory activity and amelioration of TGF-beta1-mediated fibrosis.

Kitani A, Fuss I, Nakamura K, Kumaki F, Usui T, Strober W - J. Exp. Med. (2003)

Effect of intranasal pTet-On-TGF-β1 administration on TNBS-induced colitis. (a) Weight changes of mice groups administered intrarectal TNBS and intranasal pTet-On-TGF-β or pTet-On-mock (control plasmid) with or without Dox (i.p.) Each group, n = 10. (b) LP cells from mice 5 d after TNBS colitis induction were stimulated and cultured for ELISA assays of TGF-β1, IFN-γ, IL-10, and IL-12. As shown, LP cells from mice administered Dox plus pTet-On-TGF-β1 produced large amounts of TGF-β1 and IL-10 and exhibited suppressed IL-12 and IFN-γ production. Data shown is representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2194234&req=5

fig2: Effect of intranasal pTet-On-TGF-β1 administration on TNBS-induced colitis. (a) Weight changes of mice groups administered intrarectal TNBS and intranasal pTet-On-TGF-β or pTet-On-mock (control plasmid) with or without Dox (i.p.) Each group, n = 10. (b) LP cells from mice 5 d after TNBS colitis induction were stimulated and cultured for ELISA assays of TGF-β1, IFN-γ, IL-10, and IL-12. As shown, LP cells from mice administered Dox plus pTet-On-TGF-β1 produced large amounts of TGF-β1 and IL-10 and exhibited suppressed IL-12 and IFN-γ production. Data shown is representative of three independent experiments.
Mentions: In a further study we investigated the ability of the Tet-On-TGF-β1 plasmid to affect an inflammatory state, exemplified by the Th1-type colitis induced by TNBS (4–7). Four groups of mice were studied: those receiving pTet-On-TGF-β1 with or without Dox and those receiving control plasmid (pTet-On-mock) also with or without Dox. As shown in Fig. 2 a, using body weight as an indicator of colitis, it was clear that the group that received pTet-On-TGF-β1 (i.n.) and Dox (i.p.) was protected from developing colitis, whereas the group receiving the plasmid in the absence of Dox (or those mice receiving pTet-ON-mock again with and without Dox) developed colitis. In addition, although LP cells from all three groups developing colitis produced large amounts of IL-12 and IFN-γ and small amounts of TGF-β1, LP cells from mice prevented from developing colitis produced small amounts of IL-12 and IFN-γ and very large amounts of TGF-β1 (Fig. 2 b). Of interest, the latter was much greater than that produced by cells taken from mice without inflammation, reflecting the activation of the CMV promoter in an inflammatory milieu. In addition, LP cells from this group, but not those from the groups in which the plasmid was not accompanied by Dox administration, produced large amounts of IL-10. Thus, it was obvious that such IL-10 secretion was due to plasmid-induced TGF-β1 and not simply to the presence of a Th1 inflammation. In further studies not shown, we found that intranasal administration of pTet-On-TGF-β1 also reversed TNBS colitis after inflammation was established, but only in the presence of concomitant Dox administration.

Bottom Line: In addition, using a self-inactivating retrovirus luciferase reporter construct we showed that TGF-beta1 induces Smad4, which then binds to and activates the IL-10 promoter.Furthermore, intranasal TGF-beta1 plasmid administration ameliorates bleomycin-induced fibrosis in wild-type but not IL-10-deficient mice, strongly suggesting that the amelioration is IL-10 dependent and that IL-10 protects mice from TGF-beta1-mediated fibrosis.Taken together, these findings suggest that the induction of IL-10 by TGF-beta1 is not fortuitous, but instead fulfills important requirements of TGF-beta1 function after its secretion by regulatory T cells.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 10, Room 11N238, 10 Center Drive, Bethesda, MD 20892, USA.

ABSTRACT
Interleukin (IL)-10 and transforming growth factor (TGF)-beta1 are suppressor cytokines that frequently occur together during a regulatory T cell response. Here we used a one gene doxycycline (Dox)-inducible plasmid encoding TGF-beta1 to analyze this association and test its utility. In initial studies, we showed that intranasal administration of this plasmid (along with Dox) led to the appearance of TGF-beta1-producing cells (in spleen and lamina propria) and the almost concomitant appearance of IL-10-producing cells. Moreover, we showed that these cells exert Dox-regulated suppression of the T helper cell (Th)1-mediated inflammation in trinitrobenzene sulfonic acid colitis. In subsequent in vitro studies using retroviral TGF-beta1 expression, we established that IL-10 production by Th1 cells occurs after exposure to TGF-beta1 from either an endogenous or exogenous source. In addition, using a self-inactivating retrovirus luciferase reporter construct we showed that TGF-beta1 induces Smad4, which then binds to and activates the IL-10 promoter. Furthermore, intranasal TGF-beta1 plasmid administration ameliorates bleomycin-induced fibrosis in wild-type but not IL-10-deficient mice, strongly suggesting that the amelioration is IL-10 dependent and that IL-10 protects mice from TGF-beta1-mediated fibrosis. Taken together, these findings suggest that the induction of IL-10 by TGF-beta1 is not fortuitous, but instead fulfills important requirements of TGF-beta1 function after its secretion by regulatory T cells.

Show MeSH
Related in: MedlinePlus