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Progranulin facilitates conversion and function of regulatory T cells under inflammatory conditions.

Wei F, Zhang Y, Zhao W, Yu X, Liu CJ - PLoS ONE (2014)

Bottom Line: PGRN was required for the immunosuppressive function of Tregs, since PGRN-deficient Tregs have a significant decreased ability to suppress the proliferation of effector T cells (Teff).In addition, PGRN deficiency caused a marked reduction in Tregs number in the course of inflammatory arthritis, although no significant difference was observed in the numbers of Tregs between wild type and PGRN deficient mice during development.Furthermore, PGRN deficiency led to significant upregulation of the Wnt receptor gene Fzd2.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, New York University Medical Center, New York, New York, United States of America; Institute of Pathogenic Biology, Shandong University School of Medicine, Jinan, China.

ABSTRACT
The progranulin (PGRN) is known to protect regulatory T cells (Tregs) from a negative regulation by TNF-α, and its levels are elevated in various kinds of autoimmune diseases. Whether PGRN directly regulates the conversion of CD4+CD25-T cells into Foxp3-expressing regulatory T cells (iTreg), and whether PGRN affects the immunosuppressive function of Tregs, however, remain unknown. In this study we provide evidences demonstrating that PGRN is able to stimulate the conversion of CD4+CD25-T cells into iTreg in a dose-dependent manner in vitro. In addition, PGRN showed synergistic effects with TGF-β1 on the induction of iTreg. PGRN was required for the immunosuppressive function of Tregs, since PGRN-deficient Tregs have a significant decreased ability to suppress the proliferation of effector T cells (Teff). In addition, PGRN deficiency caused a marked reduction in Tregs number in the course of inflammatory arthritis, although no significant difference was observed in the numbers of Tregs between wild type and PGRN deficient mice during development. Furthermore, PGRN deficiency led to significant upregulation of the Wnt receptor gene Fzd2. Collectively, this study reveals that PGRN directly regulates the numbers and function of Tregs under inflammatory conditions, and provides new insight into the immune regulatory mechanism of PGRN in the pathogenesis of inflammatory and immune-related diseases.

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Recombinant PGRN promotes the induction of inducible regulatory T cells in vitro.Naïve CD4+GFP- cells in lymphocytes of spleen from Foxp3-GFP reporter mice were purified by using Mitenyi reagents and a MACS apparatus. The purity of cells was evaluated by FACS method. CD4+GFP- cells conversion assay was performed as described in Methods, and GFP expression in TGF-β-unstimulated CD4+GFP- cells was taken as a control. After 3-4 days, cells were washed and GFP expression was analyzed by FACS. Data represent three independent experiments are shown. (A) No TGF-β and no PGRN. (B) No TGF-β plus 200 ng/ml of PGRN. (C) No TGF-β plus 1 µg/ml of PGRN. (D) 0.1 ng/ml of TGF-β and no PGRN. (E) 0.1 ng/ml of TGF-βplus 200 ng/ml of PGRN. (F) 0.1 ng/ml of TGF-β plus 1 µg/ml of PGRN. Data represent as a means ±SE of a representative experiment. *p<0.05; **p<0.01.
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pone-0112110-g004: Recombinant PGRN promotes the induction of inducible regulatory T cells in vitro.Naïve CD4+GFP- cells in lymphocytes of spleen from Foxp3-GFP reporter mice were purified by using Mitenyi reagents and a MACS apparatus. The purity of cells was evaluated by FACS method. CD4+GFP- cells conversion assay was performed as described in Methods, and GFP expression in TGF-β-unstimulated CD4+GFP- cells was taken as a control. After 3-4 days, cells were washed and GFP expression was analyzed by FACS. Data represent three independent experiments are shown. (A) No TGF-β and no PGRN. (B) No TGF-β plus 200 ng/ml of PGRN. (C) No TGF-β plus 1 µg/ml of PGRN. (D) 0.1 ng/ml of TGF-β and no PGRN. (E) 0.1 ng/ml of TGF-βplus 200 ng/ml of PGRN. (F) 0.1 ng/ml of TGF-β plus 1 µg/ml of PGRN. Data represent as a means ±SE of a representative experiment. *p<0.05; **p<0.01.

Mentions: We further performed the gain-of-function experiment, we treated CD4+GFP- T cells with different concentrations of PGRN and examined the change of GFP expression in CD4+ T cells. As shown in Fig. 4, PGRN significantly promoted the conversion of CD4+GFP- T cells into CD4+GFP+ T cells at a dose-dependent manner in the presence of 0.1 ng/ml TGF-β (2.74±0.70% GFP+ cells with no PGRN versus 4.33±0.98% GFP+ cells with 200 ng/ml PGRN versus 12.3±2.85% GFP+ cells with 1 µg/ml PGRN, p <0.05) (Fig. 4D–F). Furthermore, in the absence of TGF-β, 1 µg/ml of PGRN significantly induced the expression of GFP in 5.67±1.65% of the cells, when compared with 0.16±0.07% GFP+ cells without PGRN (p <0.01) (Fig. 4A and C). And low concentrations of PGRN also significantly induced GFP expression in CD4+ cells, when compared with no PGRN conditions (0.63±0.23% GFP+ cells versus 0.16±0.07% GFP+ cells, p <0.05) (Fig. 4B). Taken together, these results indicate that recombinant PGRN promotes and synergistically enhances TGF-β-mediated induction of inducible regulatory T cells in vitro.


Progranulin facilitates conversion and function of regulatory T cells under inflammatory conditions.

Wei F, Zhang Y, Zhao W, Yu X, Liu CJ - PLoS ONE (2014)

Recombinant PGRN promotes the induction of inducible regulatory T cells in vitro.Naïve CD4+GFP- cells in lymphocytes of spleen from Foxp3-GFP reporter mice were purified by using Mitenyi reagents and a MACS apparatus. The purity of cells was evaluated by FACS method. CD4+GFP- cells conversion assay was performed as described in Methods, and GFP expression in TGF-β-unstimulated CD4+GFP- cells was taken as a control. After 3-4 days, cells were washed and GFP expression was analyzed by FACS. Data represent three independent experiments are shown. (A) No TGF-β and no PGRN. (B) No TGF-β plus 200 ng/ml of PGRN. (C) No TGF-β plus 1 µg/ml of PGRN. (D) 0.1 ng/ml of TGF-β and no PGRN. (E) 0.1 ng/ml of TGF-βplus 200 ng/ml of PGRN. (F) 0.1 ng/ml of TGF-β plus 1 µg/ml of PGRN. Data represent as a means ±SE of a representative experiment. *p<0.05; **p<0.01.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4230946&req=5

pone-0112110-g004: Recombinant PGRN promotes the induction of inducible regulatory T cells in vitro.Naïve CD4+GFP- cells in lymphocytes of spleen from Foxp3-GFP reporter mice were purified by using Mitenyi reagents and a MACS apparatus. The purity of cells was evaluated by FACS method. CD4+GFP- cells conversion assay was performed as described in Methods, and GFP expression in TGF-β-unstimulated CD4+GFP- cells was taken as a control. After 3-4 days, cells were washed and GFP expression was analyzed by FACS. Data represent three independent experiments are shown. (A) No TGF-β and no PGRN. (B) No TGF-β plus 200 ng/ml of PGRN. (C) No TGF-β plus 1 µg/ml of PGRN. (D) 0.1 ng/ml of TGF-β and no PGRN. (E) 0.1 ng/ml of TGF-βplus 200 ng/ml of PGRN. (F) 0.1 ng/ml of TGF-β plus 1 µg/ml of PGRN. Data represent as a means ±SE of a representative experiment. *p<0.05; **p<0.01.
Mentions: We further performed the gain-of-function experiment, we treated CD4+GFP- T cells with different concentrations of PGRN and examined the change of GFP expression in CD4+ T cells. As shown in Fig. 4, PGRN significantly promoted the conversion of CD4+GFP- T cells into CD4+GFP+ T cells at a dose-dependent manner in the presence of 0.1 ng/ml TGF-β (2.74±0.70% GFP+ cells with no PGRN versus 4.33±0.98% GFP+ cells with 200 ng/ml PGRN versus 12.3±2.85% GFP+ cells with 1 µg/ml PGRN, p <0.05) (Fig. 4D–F). Furthermore, in the absence of TGF-β, 1 µg/ml of PGRN significantly induced the expression of GFP in 5.67±1.65% of the cells, when compared with 0.16±0.07% GFP+ cells without PGRN (p <0.01) (Fig. 4A and C). And low concentrations of PGRN also significantly induced GFP expression in CD4+ cells, when compared with no PGRN conditions (0.63±0.23% GFP+ cells versus 0.16±0.07% GFP+ cells, p <0.05) (Fig. 4B). Taken together, these results indicate that recombinant PGRN promotes and synergistically enhances TGF-β-mediated induction of inducible regulatory T cells in vitro.

Bottom Line: PGRN was required for the immunosuppressive function of Tregs, since PGRN-deficient Tregs have a significant decreased ability to suppress the proliferation of effector T cells (Teff).In addition, PGRN deficiency caused a marked reduction in Tregs number in the course of inflammatory arthritis, although no significant difference was observed in the numbers of Tregs between wild type and PGRN deficient mice during development.Furthermore, PGRN deficiency led to significant upregulation of the Wnt receptor gene Fzd2.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, New York University Medical Center, New York, New York, United States of America; Institute of Pathogenic Biology, Shandong University School of Medicine, Jinan, China.

ABSTRACT
The progranulin (PGRN) is known to protect regulatory T cells (Tregs) from a negative regulation by TNF-α, and its levels are elevated in various kinds of autoimmune diseases. Whether PGRN directly regulates the conversion of CD4+CD25-T cells into Foxp3-expressing regulatory T cells (iTreg), and whether PGRN affects the immunosuppressive function of Tregs, however, remain unknown. In this study we provide evidences demonstrating that PGRN is able to stimulate the conversion of CD4+CD25-T cells into iTreg in a dose-dependent manner in vitro. In addition, PGRN showed synergistic effects with TGF-β1 on the induction of iTreg. PGRN was required for the immunosuppressive function of Tregs, since PGRN-deficient Tregs have a significant decreased ability to suppress the proliferation of effector T cells (Teff). In addition, PGRN deficiency caused a marked reduction in Tregs number in the course of inflammatory arthritis, although no significant difference was observed in the numbers of Tregs between wild type and PGRN deficient mice during development. Furthermore, PGRN deficiency led to significant upregulation of the Wnt receptor gene Fzd2. Collectively, this study reveals that PGRN directly regulates the numbers and function of Tregs under inflammatory conditions, and provides new insight into the immune regulatory mechanism of PGRN in the pathogenesis of inflammatory and immune-related diseases.

Show MeSH
Related in: MedlinePlus