Effect of inflammatory environment on equine bone marrow derived mesenchymal stem cells immunogenicity and immunomodulatory properties
Effect of inflammatory environment on equine bone marrow derived mesenchymal stem cells immunogenicity and immunomodulatory properties
复制标题
DOI：
10.1016/j.vetimm.2016.02.007
复制
发表时间：
2016-03-01
影响因子：
1.8
通讯作者：
Rodellar, C.
中科院分区：
文献类型：
--
作者：
Barrachina, L.;Remacha, A. R.;Rodellar, C.
间充质干细胞（MSCs）由于其再生潜力，正在被研究用于马关节疾病的治疗。近年来，研究的重点主要集中在它们的免疫调节能力上。炎症在关节病理中起着核心作用，因为促炎介质释放到滑液（SF）导致软骨酶降解的激活。MSCs可以通过与免疫细胞的直接或旁分泌相互作用来调节局部免疫环境，抑制其增殖并重新定位其功能。促炎分子可诱导间充质干细胞的免疫调节潜能，但也可增加免疫原性分子的表达。研究炎症环境对间充质干细胞免疫调节和免疫原性的影响是改善细胞治疗的必要条件。本研究的目的是分析马骨髓间充质干细胞（eBM-MSCs）对三种炎症条件的反应。将来自3只动物的马BM-MSCs暴露于：(a) 20%异体炎性SF (SF)；(b) 50 ng/ml TNF α和IFN γ (CK50)和(c) 20 ng/ml TNF α和IFN γ (CK20)。暴露72 h后，通过RT-qPCR和流式细胞术分析免疫原性和免疫调节相关分子的表达，包括细胞间接触和旁分泌信号分子。在所有炎症条件下，粘附分子的基因表达上调，而MSC迁移相关基因表达下调。CK培养条件显著上调COX-2、iNOS、IDO和IL-6的表达。所有条件下MHC-I基因表达均上调，而MHC-II基因表达仅在CK启动后上调。CD40的表达没有显著变化，而配体CD40L在CK条件下下调。流式细胞术显示，在CK50条件下，MHC-I和MHC-II分子的阳性细胞百分比和平均荧光强度（MFI）增加，支持基因表达结果。这些结果加强了eBM-MSCs根据周围条件的免疫表型变化。炎症性滑膜环境没有导致显著的变化，因此eBM-MSCs在关节内给药时发现的环境可能不足以激活其免疫调节潜能。在测试剂量下，CK启动增强eBM-MSCs的免疫调节谱，这可能促进治疗效益。即使CK启动诱导MHC表达上调，但共刺激分子表达并未上调，提示免疫原性可能没有增加。这项研究提供了对炎症关节内eBM-MSCs行为的更好理解，并构成了改善基于mmsc的马关节疾病治疗的第一步。(C) 2016 Elsevier B.V.版权所有
Mesenchymal stem cells (MSCs) are being investigated for the treatment of equine joint diseases because of their regenerative potential. Recently, the focus mainly has addressed to their immunomodulatory capacities. Inflammation plays a central role in joint pathologies, since the release of proinflammatory mediators to the synovial fluid (SF) leads to the activation of enzymatic degradation of the cartilage. MSCs can modulate the local immune environment through direct or paracrine interaction with immune cells, suppressing their proliferation and re-addressing their functions. Proinflammatory molecules can induce MSC immunoregulatory potential, but they could also increase the expression of immunogenic molecules. Studying the effect of inflammatory environment on MSC immunomodulation and immunogenicity profiles is mandatory to improve cellular therapies. The aim of this study was to analyse the response of equine bone marrow MSCs (eBM-MSCs) to three inflammatory conditions. Equine BM-MSCs from three animals were exposed to: (a) 20% allogeneic inflammatory SF (SF); (b) 50 ng/ml of TNF alpha and IFN gamma (CK50) and (c) 20 ng/ml of TNF alpha and IFN gamma (CK20). After 72 h of exposure, expression of immunogenic and immunomodulation-related molecules, including cell-to-cell contact and paracrine signalling molecules, were analysed by RT-qPCR and flow cytometry. The gene expression of adhesion molecules was upregulated whereas MSC migration-related genes were downregulated by all inflammatory conditions tested. CK culture conditions significantly upregulated the expression of COX-2, iNOS, IDO and IL-6. MHC-I gene expression was upregulated by all conditions, whereas MHC-II was upregulated only after CK priming. The expression of CD40 did not significantly change, whereas the ligand, CD40L, was downregulated in CK conditions. Flow cytometry showed an increase in the percentage of positive cells and mean fluorescence intensity (MFI) of the MHC-I and MHC-II molecules at CK50 conditions, supporting the gene expression results. These outcomes reinforce the change of the immunophenotype of the eBM-MSCs according to the surrounding conditions. Inflammatory synovial environment did not lead to significant changes, so the environment found by eBM-MSCs when they are intraarticular administered may not be enough to activate their immunomodulatory potential. CK priming at tested doses enhances the immunoregulatory profile of eBM-MSCs, which may promote a therapeutic benefit. Even if CK priming induced an upregulation of MHC expression, costimulatory molecule expression however was not upregulated, suggesting that immunogenicity might not be increased. This study provides a better understanding about the behaviour of eBM-MSCs inside the inflamed joint and constitutes a first step to improve MSC-based therapies for equine joint diseases. (C) 2016 Elsevier B.V. All rights reserved.