间充质干细胞来源的外泌体对诱导小鼠耐受树突状细胞的影响。
Effect of mesenchymal stem cell-derived exosomes on the induction of mouse tolerogenic dendritic cells.
机构信息
Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Mycobacteriology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
出版信息
J Cell Physiol. 2020 Oct;235(10):7043-7055. doi: 10.1002/jcp.29601. Epub 2020 Feb 11.
Dendritic cells (DCs) orchestrate innate inflammatory responses and adaptive immunity through T-cell activation via direct cell-cell interactions and/or cytokine production. Tolerogenic DCs (tolDCs) help maintain immunological tolerance through the induction of T-cell unresponsiveness or apoptosis, and generation of regulatory T cells. Mesenchymal stromal cells (MSCs) are adult multipotent cells located within the stroma of bone marrow (BM), but they can be isolated from virtually all organs. Extracellular vesicles and exosomes are released from inflammatory cells and act as messengers enabling communication between cells. To investigate the effects of MSC-derived exosomes on the induction of mouse tolDCs, murine adipose-derived MSCs were isolated from C57BL/6 mice and exosomes isolated by ExoQuick-TC kits. BM-derived DCs (BMDCs) were prepared and cocultured with MSCs-derived exosomes (100 μg/ml) for 72 hr. Mature BMDCs were derived by adding lipopolysaccharide (LPS; 0.1μg/ml) at Day 8 for 24 hr. The study groups were divided into (a) immature DC (iDC, Ctrl), (b) iDC + exosome (Exo), (c) iDC + LPS (LPS), and (d) iDC + exosome + LPS (EXO + LPS). Expression of CD11c, CD83, CD86, CD40, and MHCII on DCs was analyzed at Day 9. DC proliferation was assessed by coculture with carboxyfluorescein succinimidyl ester-labeled BALB/C-derived splenocytes p. Interleukin-6 (IL-6), IL-10, and transforming growth factor-β (TGF-β) release were measured by enzyme-linked immunosorbent assay. MSC-derived exosomes decrease DC surface marker expression in cells treated with LPS, compared with control cells ( ≤ .05). MSC-derived exosomes decrease IL-6 release but augment IL-10 and TGF-β release (p ≤ .05). Lymphocyte proliferation was decreased (p ≤ .05) in the presence of DCs treated with MSC-derived exosomes. CMSC-derived exosomes suppress the maturation of BMDCs, suggesting that they may be important modulators of DC-induced immune responses.
树突状细胞 (DCs) 通过直接细胞间相互作用和/或细胞因子产生来激活 T 细胞,从而协调先天炎症反应和适应性免疫。耐受诱导型树突状细胞 (tolDCs) 通过诱导 T 细胞无反应性或凋亡以及产生调节性 T 细胞来帮助维持免疫耐受。间充质基质细胞 (MSCs) 是位于骨髓 (BM) 基质中的成体多能细胞,但它们可以从几乎所有器官中分离出来。细胞外囊泡和外泌体由炎症细胞释放,并作为信使,使细胞之间能够进行通讯。为了研究 MSC 衍生的外泌体对诱导小鼠 tolDCs 的影响,从 C57BL/6 小鼠中分离出小鼠脂肪来源的 MSC,并通过 ExoQuick-TC 试剂盒分离出外泌体。用脂多糖 (LPS;0.1μg/ml) 在第 8 天诱导 24 小时诱导 BM 来源的 DC (BMDCs) 成熟。将研究组分为 (a) 未成熟 DC (iDC,对照)、(b) iDC+外泌体 (Exo)、(c) iDC+LPS (LPS) 和 (d) iDC+外泌体+LPS (EXO+LPS)。第 9 天分析 DC 上 CD11c、CD83、CD86、CD40 和 MHCII 的表达。通过与羧基荧光素琥珀酰亚胺酯标记的 BALB/C 来源的脾细胞共培养来评估 DC 增殖。通过酶联免疫吸附试验测量白细胞介素 6 (IL-6)、白细胞介素 10 (IL-10) 和转化生长因子-β (TGF-β) 的释放。与对照细胞相比,用 LPS 处理的细胞中 MSC 衍生的外泌体降低了 DC 表面标志物的表达 (≤.05)。MSC 衍生的外泌体降低了 IL-6 的释放,但增加了 IL-10 和 TGF-β 的释放 (p≤.05)。在存在用 MSC 衍生的外泌体处理的 DC 的情况下,淋巴细胞增殖减少 (p≤.05)。CMSC 衍生的外泌体抑制 BMDC 的成熟,这表明它们可能是 DC 诱导的免疫反应的重要调节剂。