Introduction

Mesenchymal stem cells (MSCs) have always been the center of the experimental exploration of regenerative therapy together with other stem cells. Among with, peripheral blood-derived mesenchymal stem cells (PBMSCs) have been regarded as promising in clinical applications for its convenience of acquisition from peripheral blood. However, few reported experiments so far to elucidate the exact mechanisms of how PBMSC influence regeneration. As the ability of immunomodulatory is one of the crucial features that influence MSC to reconstruct impaired tissue, we decided to focus on the immunomodulatory abilities of PBMSCs and conducted experiments associated with macrophages and T lymphocytes, which are two main cell types that dominate the innate and acquired immunity. Therefore, a basis can be made from these experiments for applications of PBMSCs in regenerative therapy in the future.

Methods

A Transwell system was used for the coculturing of PBMSCs with macrophages. T lymphocytes were cultured directly with PBMSCs. Flow cytometry and immunochemistry were conducted for identifying the phenotypes. Immunomagnetic microspheres, ELISA and RT-qPCR were used to detect the expressions of relevant molecules or mRNAs.

Results

After coculturing PBMSCs with M0, the anti-inflammatory IL-10 was increased whereas the proinflammatory TNF-α decreased; the expression of CD11b, CD68, CD206, Arg-1, IL-10 and CCL-22 was up-regulated whereas IL-1β down-regulated. The expression of TGF-β, RORγt, Foxp3 and IL-10 was increased in the cocultured lymphocytes whereas IL-17 and IL-6 decreased; the ratio of CD4⁺IL-17⁺ Th17/CD25⁺Foxp3⁺ Treg was reduced.

Conclusion

The findings demonstrated that PBMSCs promoted the anti-inflammatory features of macrophages and the Th17/Treg system. PBMSCs are able to inhibit inflammation associated with these two immune cell systems, and thus provide insight into how PBMSCs achieve their immunomodulatory ability.

中文翻译：

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外周血间充质干细胞对体外巨噬细胞极化和Th17/Treg平衡的影响。

介绍

间充质干细胞（MSCs）一直是与其他干细胞一起进行再生治疗实验探索的中心。其中，外周血来源的间充质干细胞（PBMSCs）因其从外周血中获取的便利性而被认为在临床应用中很有前景。然而，到目前为止，很少有实验报告阐明 PBMSC 如何影响再生的确切机制。由于免疫调节能力是影响 MSC 重建受损组织的关键特征之一，我们决定专注于 PBMSCs 的免疫调节能力，并进行与巨噬细胞和 T 淋巴细胞相关的实验，这两种主要细胞类型在先天和获得性免疫。所以，

方法

Transwell 系统用于 PBMSCs 与巨噬细胞的共培养。T淋巴细胞直接与PBMSCs一起培养。进行流式细胞术和免疫化学以鉴定表型。采用免疫磁性微球、ELISA和RT-qPCR检测相关分子或mRNA的表达。

结果

PBMSCs与M0共培养后，抗炎IL-10升高，促炎TNF-α降低；CD11b、CD68、CD206、Arg-1、IL-10 和 CCL-22 的表达上调，而 IL-1β 下调。共培养的淋巴细胞中TGF-β、RORγt、Foxp3和IL-10的表达增加，而IL-17和IL-6的表达减少；CD4 ⁺ IL-17 ⁺ Th17/CD25 ⁺ Foxp3 ⁺ Treg 的比例降低。

结论

研究结果表明，PBMSCs 促进了巨噬细胞和 Th17/Treg 系统的抗炎特征。PBMSCs 能够抑制与这两种免疫细胞系统相关的炎症，从而深入了解 PBMSCs 如何实现其免疫调节能力。