Fuenzalida Patricia, Kurte Mónica, Fernández-O'ryan Catalina, Ibañez Cristina, Gauthier-Abeliuk Melanie, Vega-Letter Ana María, Gonzalez Paz, Irarrázabal Carlos, Quezada Nataly, Figueroa Fernando, Carrión Flavio
Cellular and Molecular Immunology Laboratory, Faculty of Medicine, University of the Andes, Santiago, Chile.
Laboratory of Nano-Regenerative Medicine, Faculty of Medicine, University of the Andes, Santiago, Chile; Cells for Cells, Santiago, Chile, Faculty of Medicine, University of the Andes, Santiago, Chile.
Cytotherapy. 2016 May;18(5):630-41. doi: 10.1016/j.jcyt.2016.02.002.
Immunomodulatory properties of human umbilical cord-derived mesenchymal stromal cells (UCMSCs) can be differentially modulated by toll-like receptors (TLR) agonists. Here, the therapeutic efficacy of short TLR3 and TLR4 pre-conditioning of UCMSCs was evaluated in a dextran sulfate sodium (DSS)-induced colitis in mice. The novelty of this study is that although modulation of human MSCs activity by TLRs is not a new concept, this is the first time that short TLR pre-conditioning has been carried out in a murine inflammatory model of acute colitis.
C57BL/6 mice were exposed to 2.5% dextran sulfate sodium (DSS) in drinking water ad libitum for 7 days. At days 1 and 3, mice were injected intraperitoneally with 1 × 10(6) UCMSCs untreated or TLR3 and TLR4 pre-conditioned UCMSCs. UCMSCs were pre-conditioned with poly(I:C) for TLR3 and LPS for TLR4 for 1 h at 37°C and 5% CO2. We evaluated clinical signs of disease and body weights daily. At the end of the experiment, colon length and histological changes were assessed.
poly(I:C) pre-conditioned UCMSCs significantly ameliorated the clinical and histopathological severity of DSS-induced colitis compared with UCMSCs or LPS pre-conditioned UCMSCs. In contrast, infusion of LPS pre-conditioned UCMSCs significantly increased clinical signs of disease, colon shortening and histological disease index in DSS-induced colitis.
These results show that short in vitro TLR3 pre-conditioning with poly(I:C) enhances the therapeutic efficacy of UCMSCs, which is a major breakthrough for developing improved treatments to patients with inflammatory bowel disease.
人脐带间充质基质细胞（UCMSCs）的免疫调节特性可被 Toll 样受体（TLR）激动剂差异性调节。在此，我们评估了对 UCMSCs 进行短暂 TLR3 和 TLR4 预处理在葡聚糖硫酸钠（DSS）诱导的小鼠结肠炎中的治疗效果。本研究的新颖之处在于，虽然 TLRs 对人骨髓间充质干细胞活性的调节并非新概念，但这是首次在急性结肠炎的小鼠炎症模型中进行短暂的 TLR 预处理。
C57BL/6 小鼠自由饮用含 2.5%葡聚糖硫酸钠（DSS）的水 7 天。在第 1 天和第 3 天，小鼠腹腔注射 1×10⁶ 未处理的 UCMSCs 或经 TLR3 和 TLR4 预处理的 UCMSCs。UCMSCs 分别用聚肌苷酸胞苷酸（poly(I:C)）预处理 TLR3、用脂多糖（LPS）预处理 TLR4，在 37°C 和 5%二氧化碳条件下处理 1 小时。我们每天评估疾病的临床症状和体重。实验结束时，评估结肠长度和组织学变化。
与未处理的 UCMSCs 或经 LPS 预处理的 UCMSCs 相比，经 poly(I:C)预处理的 UCMSCs 显著改善了 DSS 诱导的结肠炎的临床和组织病理学严重程度。相反，输注经 LPS 预处理的 UCMSCs 显著增加了 DSS 诱导的结肠炎的疾病临床症状、结肠缩短和组织学疾病指数。
这些结果表明，用 poly(I:C)进行短暂的体外 TLR3 预处理可增强 UCMSCs 的治疗效果，这是开发针对炎症性肠病患者的改进治疗方法的一项重大突破。
