Mesenchymal Stromal Cell-Derived Exosomes Affect mRNA Expression and Function of B-Lymphocytes
Mesenchymal Stromal Cell-Derived Exosomes Affect mRNA Expression and Function of B-Lymphocytes
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DOI：
10.3389/fimmu.2018.03053
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发表时间：
2018-12-21
影响因子：
7.3
通讯作者：
Avni, Batia
中科院分区：
文献类型：
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作者：
Khare, Drirh;Or, Reuven;Avni, Batia
背景资料：骨髓间充质干细胞（bmMSC）可能在调节淋巴细胞的成熟、增殖和功能活化方面发挥作用，但确切机制尚不清楚。MSC衍生的外泌体诱导B、T和单核细胞衍生的树突细胞功能的调节应答。在这里，我们评估了人淋巴细胞的bmMSC衍生的外泌体的特异性抑制和对B-细胞function.Methods的影响：外泌体从培养液中分离bmMSC从几个健康供体。通过羧基荧光素琥珀酰亚胺酯测定法测定纯化的bmMSC衍生的exosomes对活化的外周血单个核细胞（PBMC）和分离的B和T淋巴细胞增殖的影响。使用Illumina测序平台，对在存在或不存在外来体的情况下活化的B淋巴细胞进行mRNA谱分析。应用Insiderity（R）通路分析软件分析通路网络，并对差异表达基因的生物学功能进行分析。通过RT-PCR进行验证。结果：与对照组相比，与MSC衍生的外泌体共培养的活化PBMC或分离的T和B细胞的增殖分别降低了37%、23%和18%。激活的B淋巴细胞的mRNA谱显示了186个基因在外泌体处理的细胞和对照细胞之间差异表达。我们观察到参与细胞运输、发育、止血和免疫细胞功能的基因的下调和上调。通过真实的时间PCR分析CXCL 8（IL 8）和MZB 1基因的表达来验证RNA-Seq结果，已知CXCL 8和MZB 1基因在免疫调节中具有重要作用。IgM产生水平降低证实了功能改变。一致的结果在各种各样的健康人bmMSC donors.Conclusion：我们的数据表明，外泌体可能在免疫调节中发挥重要作用。它们抑制几种类型的免疫细胞的增殖。在B淋巴细胞中，它们通过发挥相关基因mRNA的差异表达来调节细胞功能。这项研究的结果有助于阐明外泌体诱导免疫调节的机制，并可能有助于开发更新，更安全的治疗策略。
Background: Bone marrow mesenchymal stem cells (bmMSC) may play a role in the regulation of maturation, proliferation, and functional activation of lymphocytes, though the exact mechanisms are unknown. MSC-derived exosomes induce a regulatory response in the function of B, T, and monocyte-derived dendritic cells. Here, we evaluated the specific inhibition of human lymphocytes by bmMSC-derived exosomes and the effects on B-cell function.Methods: Exosomes were isolated from culture media of bmMSC obtained from several healthy donors. The effect of purified bmMSC-derived exosomes on activated peripheral blood mononuclear cells (PBMCs) and isolated B and T lymphocyte proliferation was measured by carboxyfluorescein succinimidyl ester assay. Using the Illumina sequencing platform, mRNA profiling was performed on B-Iymphocytes activated in the presence or absence of exosomes. Ingenuity (R) pathway analysis software was applied to analyze pathway networks, and biological functions of the differentially expressed genes. Validation by RT-PCR was performed. The effect of bmMSC-derived exosomes on antibody secretion was measured by ELISA.Results: Proliferation of activated PBMCs or isolated T and B cells co-cultured with MSC-derived exosomes decreased by 37, 23, and 18%, respectively, compared to controls. mRNA profiling of activated B-Iymphocytes revealed 186 genes that were differentially expressed between exosome-treated and control cells. We observed down- and up-regulation of genes that are involved in cell trafficking, development, hemostasis, and immune cell function. RNA-Seq results were validated by real time PCR analysis for the expression of CXCL8 (IL8) and MZB1 genes that are known to have an important role in immune modulation. Functional alterations were confirmed by decreased IgM production levels. Consistent results were demonstrated among a wide variety of healthy human bmMSC donors.Conclusion: Our data show that exosomes may play an important role in immune regulation. They inhibit proliferation of several types of immune cells. In B-lymphocytes they modulate cell function by exerting differential expression of the mRNA of relevant genes. The results of this study help elucidate the mechanisms by which exosomes induce immune regulation and may contribute to the development of newer and safer therapeutic strategies.