Institute of Basic Medical Sciences and School of Basic Medicine, Center of Excellence in Tissue Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
Exp Hematol. 2011 Feb;39(2):214-224.e1. doi: 10.1016/j.exphem.2010.10.009. Epub 2010 Nov 13.
Cyclosporine A (CsA), known as an effective immunosuppressive agent, is widely used in clinical fields. Mesenchymal stem cells may exert immunomodulatory effects on the immune system, but the exact mechanisms underlying them remain controversial. Here we investigated whether human adipose tissue-derived mesenchymal stem cells (AMSCs) facilitate in vitro the immunomodulatory effects of CsA and we explored the molecule mechanisms that may be involved.
Proliferation of T lymphocytes was measured by uptake of (3)H-thymidine. Transcription and production of interleukin-2 and interferon-γ were evaluated by real-time quantitative polymerase chain reaction, reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay. Nuclear factor-κB (NF-κB) was assayed by Western blotting and electrophoretic mobility shift assay. Expression of Jagged-1, Jagged-2, and Delta-1 of AMSCs were surveyed by flow cytometric analysis and Western blotting.
The combination of moderate-dose AMSCs and low-dose CsA was significantly more powerful than moderate-dose AMSCs or large-dose CsA alone in suppressing transcription and production of interleukin-2 and interferon-γ, activation of NF-κB, and proliferation of T lymphocytes. In addition, AMSCs expressed a high level of Jagged-1, which induced activation of Notch signaling in T lymphocytes, thus reducing NF-κB activity. Anti-Jagged-1 neutralizing antibody and N [N-(3, 5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butyl ester could reverse this trend.
Human AMSCs facilitate the immunosuppressive effect of CsA on T lymphocytes through Jagged-1/Notch-related inhibition of NF-κB signaling. The combination of AMSCs and CsA represents a rationale therapeutic approach aimed to prevent adverse effects of CsA while maintaining its adequate immunosuppressive effect. Expression of Jagged-1 on AMSCs may provide an effective mechanism for the immunomodulatory activity of AMSCs via direct cell-cell interaction.
环孢素 A（CsA）作为一种有效的免疫抑制剂，在临床领域得到广泛应用。间充质干细胞（MSCs）可能对免疫系统发挥免疫调节作用，但确切的机制仍存在争议。本研究旨在探讨人脂肪组织来源的间充质干细胞（AMSCs）是否有助于 CsA 的体外免疫调节作用，并探讨可能涉及的分子机制。
通过摄取[3]H-胸苷来测量 T 淋巴细胞的增殖。通过实时定量聚合酶链反应、逆转录聚合酶链反应和酶联免疫吸附试验评估白细胞介素-2 和干扰素-γ的转录和产生。通过 Western 印迹和电泳迁移率变动分析测定核因子-κB（NF-κB）。通过流式细胞术分析和 Western 印迹检测 AMSCs 的 Jagged-1、Jagged-2 和 Delta-1 的表达。
与单独使用中剂量 AMSCs 或大剂量 CsA 相比，中剂量 AMSCs 与低剂量 CsA 的联合使用在抑制白细胞介素-2 和干扰素-γ的转录和产生、NF-κB 的激活以及 T 淋巴细胞的增殖方面更为有效。此外，AMSCs 表达高水平的 Jagged-1，诱导 T 淋巴细胞 Notch 信号的激活，从而降低 NF-κB 的活性。抗 Jagged-1 中和抗体和 N-[N-（3，5-二氟苯乙酰基-L-丙氨酰）]-S-苯甘氨酸叔丁酯可以逆转这一趋势。
人 AMSCs 通过 Jagged-1/Notch 相关抑制 NF-κB 信号通路促进 CsA 对 T 淋巴细胞的免疫抑制作用。AMSCs 与 CsA 的联合应用为预防 CsA 的不良反应同时保持其适当的免疫抑制作用提供了一种合理的治疗方法。AMSCs 上 Jagged-1 的表达可能通过直接的细胞-细胞相互作用为 AMSCs 的免疫调节活性提供有效的机制。
