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一种鉴定具有高免疫抑制潜能的骨髓间充质基质细胞的简单方法。

A simple method for identifying bone marrow mesenchymal stromal cells with a high immunosuppressive potential.

机构信息

Department of Experimental and Diagnostic Medicine, Laboratory of Immunogenetics, Section of Medical Genetics, University of Ferrara, Ferrara, Italy.

出版信息

Cytotherapy. 2011 May;13(5):523-7. doi: 10.3109/14653249.2010.542460. Epub 2010 Dec 20.

Abstract

BACKGROUND AIMS

The beneficial activity of mesenchymal stromal cells (MSC) in allogeneic hematopietic stem cell transplantation requires correct use in terms of cell dose and timing of infusion and the identification of biomarkers for selection. The immunosuppressive bone marrow (BM)-derived MSC (BM-MSC) functions have been associated with the production of soluble HLA-G molecules (sHLA-G) via interleukin (IL)-10. We have established a reliable method for evaluating BM-MSC HLA-G expression without the influence of peripheral blood mononuclear cells (PBMC).

METHODS

Thirteen BM-MSC from donors were activated with recombinant IL-10 or co-cultured with 10 different phytohemagglutinin (PHA)-treated PBMC (PHA-PBMC). Membrane-bound and sHLA-G expression was evaluated by flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively; lymphoproliferation was measured by (methyl-(3)H)thymidine.

RESULTS

The results demonstrated the ability of IL-10 to stimulate both membrane-bound and sHLA-G production by BM-MSC. The levels of HLA-G expression induced by IL-10 in BM-MSC were associated with the inhibition of PHA-PBMC proliferation (sHLA-G, P = 0.0008, r = 0.9308; membrane HLA-G, P = 0.0005, r = 0.9502).

CONCLUSIONS

We propose the evaluation of sHLA-G production in IL-10-treated BM-MSC cultures as a possible marker of immunoregulatory function.

摘要

背景目的

间充质基质细胞(MSC)在异基因造血干细胞移植中的有益活性需要在细胞剂量和输注时间方面正确使用,并确定选择的生物标志物。骨髓(BM)来源的免疫抑制MSC(BM-MSC)功能与通过白细胞介素(IL)-10产生可溶性 HLA-G 分子(sHLA-G)有关。我们已经建立了一种可靠的方法来评估 BM-MSC HLA-G 表达,而不受外周血单核细胞(PBMC)的影响。

方法

用重组 IL-10 激活 13 名供体的 BM-MSC,或与 10 种不同的植物血球凝集素(PHA)处理的 PBMC(PHA-PBMC)共培养。通过流式细胞术和酶联免疫吸附试验(ELISA)分别评估膜结合和 sHLA-G 表达;通过(甲基-(3)H)胸腺嘧啶测量淋巴细胞增殖。

结果

结果表明,IL-10 能够刺激 BM-MSC 产生膜结合和 sHLA-G。IL-10 在 BM-MSC 中诱导的 HLA-G 表达水平与 PHA-PBMC 增殖的抑制相关(sHLA-G,P = 0.0008,r = 0.9308;膜 HLA-G,P = 0.0005,r = 0.9502)。

结论

我们提出评估 IL-10 处理的 BM-MSC 培养物中 sHLA-G 的产生作为免疫调节功能的可能标志物。

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