9 research outputs found

    MDS-MSC inhibit T-lymphocyte proliferation.

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    <p>Irradiated (15 Gy) MDS-MSC or normal-MSC were cultured for 5 days with CD2+ T-lymphocyte in the presence of PHA, then assessed by [3H]-thymidine incorporation. Data are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05.</p

    Effect of MDS-MSC on T cell apoptosis.

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    <p>T cells were incubated for 3 days alone or with MDS-MSC or normal-MSC in the presence of the mitogen PHA.The test was conducted by Annexin-V and PI double staining and analyzed by flow cytometry. Data are expressed as mean±SD of triplicates of 5 separate experiments. Annexin V+ means the cells were PI negative and Annexin V positive. *P≤0.05.</p

    Induction of CD4+CD25+Foxp3+Tregs by MDS-MSC is dependent on TGFβ1.

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    <p>Western blot confirmed efficient knockdown of TGF-β1. (B) CD4+CD25-T cells were cultured with TGF-β1 knockdown MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05. (C) CD4+CD25-T cells were cultured with mutant siRNA transfected MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 6 separate experiments. *P≤0.05. (D) anti-rhTGF-β1 mAb was added at the beginning of coculture of CD4+CD25-T cells and untransfected MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 6 separate experiments. *P≤0.05.</p

    MDS-MSC induce CD4+CD25+Foxp3+Tregs.

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    <p>(A) CD4+CD25-T cells were cultured with MDS-MSC or normal-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 4 separate experiments. *P≤0.05. (B) CD4+T cells were cocultured with MDS-MSC generated CD4+CD25+Foxp3+Tregs or normal-MSC generated CD4+CD25+Foxp3+Tregs in the presence of PHA, and the T-lymphocyte proliferation was measured on day 5 by [3H]-thymidine incorporation. Results are expressed as mean±SD of triplicates of 4 separate experiments. *P≤0.05. (C) MDS-MSC generated CD4+CD25+Foxp3+Tregs or normal-MSC generated CD4+CD25+Foxp3+Tregs inhibited the response of allogeneic T-lymphocyte in a dose-dependent manner. Responder CD2+ T-lymphocyte were stimulated with PHA for 5 days with or without graded dosed of MDS-MSC generated CD4+CD25+Foxp3+Tregs or normal-MSC generated CD4+CD25+Foxp3+Tregs. Results are expressed as mean±SD of triplicates of 4 separate experiments. *p≤0.05. (D) CD4+CD25-T cells were cultured with high-risk MDS-MSC or low-risk MDS-MSC for 5 days, and CD4+ T cells were collected. The expression of CD25 and Foxp3 on CD4+ T cells was analyzed by FACS. Results are expressed as mean±SD of triplicates of 4 separate experiments. *P≤0.05.</p

    RRs and 95% CIs for (A) 2-year relapse risk and (B) 5-year OS, according to a subgroup analysis of CBF-AML.

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    <p>The number of included studies, number of patients in the included studies, and percentage of patients with <i>KIT</i> mutations in the included studies are listed.</p

    MDS-MSC inhibit the endocytosis of monocyte derived DCs.

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    <p>DCs <b>(5×10<sup>5</sup>)</b> were incubated in medium with FITC-labeled dextran at a concentration of 1 mg/ml in the presence of MSC <b>(5×10<sup>4</sup>)</b> at day 7. After an incubation period of 60 min at 37°C (green line) or 4°C as a control (gray line), cells were harvested and analyzed by FACS.Numbers in histograms indicate the mean fluorescence of each cell population. Results are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05.</p

    MDS-MSC reduce IL-12 secretion of monocyte-derived DCs.

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    <p>DCs <b>(1×10<sup>6</sup>)</b> obtained from monocytes after 7 days of induction with GM-CSF plus IL-4 were stimulated by LPS for an additional 48 hours with or without MSC coculture, and then cell-free supernatants were collected and quantified by ELISA for IL-12 production. Results are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05.</p

    TGFβ1 plays a key role in the MSC-mediated inhibition of DCs differentiation and functions.

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    <p>Monocytes were cultured with GM-CSF and IL-4 to induce differentiation into DCs. Cultures were performed either in the absence or in the presence of MSC. In addition, anti-rhTGF-β1, or anti-rhIL-6, or 1-methyltryptophan, IDO inhibitor, or indomethacin, PGE2 inhibitor, or PBIT, NO inhibitor was added to monocyte-MSC cocultures. After 5 days, expression of CD1a (A) and CD14 (B) in cells cultured under the described was performed to check DCs differentiation. (C) IL-12 was measured in culture supernatants after 48-hour stimulation with LPS of monocyte-derived cells cultured for 7 days with GM-CSF and IL-4 either in the absence or in the presence of MSC., anti-rhTGF-β1, or anti-rhIL-6, or 1-methyltryptophan, IDO inhibitor, or indomethacin, PGE2 inhibitor, or PBIT, NO inhibitor was added at the beginning of the culture. DCs obtained from monocytes after 7 days of induction with GM-CSF plus IL-4 were stimulated by LPS for an additional 48 hours without MSC coculture. (D) anti-rhTGF-β1, or anti-rhIL-6, or 1-methyltryptophan, IDO inhibitor, or indomethacin, PGE2 inhibitor, or PBIT, NO inhibitor was added at the beginning of MLR coculture. T-lymphocyte proliferation was assessed by [3H]-thymidine incorporation. Data are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05.</p

    MDS-MSC inhibit the capability of inducing T-cell response in MLR of monocyte-derived DCs.

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    <p>mDCs <b>(1×10<sup>5</sup>)</b> were irradiated and used to induce allogeneic T cells (10<sup>6</sup> responder cells/well) with or without MSC <b>(1×10<sup>5</sup>)</b> in the MLR culture. After culture for 5 days, T-cell proliferation was evaluated by adding 3H-thymidine to each well 18 h before the cultures were terminated. Results are expressed as mean±SD of triplicates of 5 separate experiments. *P≤0.05.</p
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