Laboratory of Translational Immunology, Division of Immunology and Allergology, Department of Medical Specialties, Medical Faculty, Geneva University Hospitals, Geneva, Switzerland.
Department of Surgery, Medical Faculty, Cell Isolation and Transplantation Center, Geneva University Hospitals, Geneva, Switzerland.
Xenotransplantation. 2020 Jul;27(4):e12584. doi: 10.1111/xen.12584. Epub 2020 Jan 26.
Tolerogenic dendritic cells (DCs) represent a promising approach to promote transplantation tolerance. In this study, the potential of autologous bone marrow (BM)-derived murine DC to protect rat-to-mouse islets xenografts was analyzed.
Tolerogenic DCs were generated by differentiating BM cells in the presence of granulocyte-macrophage colony-stimulating factor and interleukin 10 (IL-10, IL-10 DC). The phenotype of IL-10 DC was characterized in vitro by expression of costimulatory/inhibitory molecules (flow cytometry) and cytokines (Luminex and ELISA), their function by phagocytosis and T-cell stimulation assays. To study transplant tolerance in vivo, rat islets were transplanted alone or in combination with autologous murine IL-10 DC under the kidney capsule of streptozotocin-induced diabetic C57BL/6 mice. Xenograft survival was evaluated by monitoring glycemia, cellular infiltration of xenografts by microscopy and flow cytometry 10 days post-transplantation.
Compared with control DC, IL-10 DC exhibited lower levels of major histocompatibility complex class II, costimulatory molecules (CD40, CD86, CD205), lower production of pro-inflammatory cytokines (IL-12p70, TNF, IL-6), and higher production of IL-10. Phagocytosis of xenogeneic rat splenocytes was not impaired in IL-10 DC, whereas stimulation of T-cell proliferation was reduced in the presence of IL-10 DC. Xenograft survival of rat islets in diabetic mice co-transplanted with autologous murine IL-10 DC was significantly prolonged from 12 to 21 days, without additional immunosuppressive treatment. Overall, infiltration of xenografts by T cells and myeloid cells was not different in IL-10 DC recipient mice, but enriched for CD8 T cells and myeloid cells with suppressor-associated phenotype.
Autologous IL-10-differentiated DC with tolerogenic properties prolong rat-to-mouse islets xenograft survival, potentially by locally inducing immune regulatory cells, indicating their potential for regulatory immune cell therapy in xenotransplantation.
诱导免疫耐受的树突状细胞（DC）代表了促进移植耐受的一种很有前途的方法。在这项研究中，分析了自体骨髓（BM）来源的小鼠 DC 保护大鼠到小鼠胰岛异种移植物的潜力。
通过在粒细胞-巨噬细胞集落刺激因子和白细胞介素 10（IL-10）存在下分化 BM 细胞来产生耐受原性 DC（IL-10 DC）。通过流式细胞术检测共刺激/抑制分子（流式细胞术）和细胞因子（Luminex 和 ELISA）的表达，吞噬作用和 T 细胞刺激试验来评估 IL-10 DC 的功能，来鉴定 IL-10 DC 的表型。为了研究体内移植耐受，将大鼠胰岛单独或与自体鼠 IL-10 DC 一起在链脲佐菌素诱导的糖尿病 C57BL/6 小鼠的肾脏囊下移植。通过监测血糖、移植后 10 天通过显微镜和流式细胞术观察异种移植物的细胞浸润，来评估异种移植物的存活。
与对照 DC 相比，IL-10 DC 表达较低水平的主要组织相容性复合体 II、共刺激分子（CD40、CD86、CD205）、较低水平的促炎细胞因子（IL-12p70、TNF、IL-6）和较高水平的 IL-10。IL-10 DC 对异种大鼠脾细胞的吞噬作用没有受损，而在存在 IL-10 DC 的情况下刺激 T 细胞增殖减少。与未接受免疫抑制治疗的糖尿病小鼠相比，自体鼠 IL-10 DC 共移植可显著延长大鼠胰岛异种移植物的存活时间，从 12 天延长至 21 天。在 IL-10 DC 受者小鼠中，异种移植物的浸润 T 细胞和髓样细胞没有差异，但富含具有抑制相关表型的 CD8 T 细胞和髓样细胞。
具有免疫耐受特性的自体 IL-10 分化的 DC 可延长大鼠到小鼠胰岛异种移植物的存活时间，可能是通过局部诱导免疫调节细胞，表明其在异种移植中的调节免疫细胞治疗的潜力。
