Department of Kidney Transplantation, Hospital of Nephropathy, The First Affiliated Hospital of Medical College of Xi'an Jiaotong University, 710061 Xi'an, Shaanxi, China.
Institute of Organ Transplantation, Xi'an Jiaotong University, 710061 Xi'an, Shaanxi, China.
Front Biosci (Landmark Ed). 2023 May 11;28(5):91. doi: 10.31083/j.fbl2805091.
The use of immature dendritic cells (imDCs) to induce donor-specific immunotolerance following stimulation is limited by their low rate of induction and their tendency to undergo maturation. We derived imDCs from bone marrow hematopoietic stem cells (HSCs-imDCs). We then tested the ability of naringenin (Nar) to impede the maturation of HSCs-imDCs for inducing transplantation immune tolerance.
HSCs derived from bone marrow were collected and induced to differentiate into imDCs by treating with Nar (Nar-HSCs-imDCs). Flow cytometry was used to evaluate DC surface markers, apoptosis, and endocytic ability. The ability of DCs to influence the proliferation of T cells and of regulatory T cells (Tregs) was analyzed by mixed lymphocyte reaction assays. Enzyme-linked immunoassays were used to quantify cytokine levels in supernatants from co-cultured DCs and Tregs, as well as in the serum of experimental animals. The level of immunotolerance induced by Nar-HSCs-imDCs was evaluated by skin grafting in recipient Balb/c mice, while the Kaplan-Meier method was used to statistically evaluate graft survival.
Compared with HSC-imDCs, Nar-HSCs-imDCs showed higher expression of cluster of differentiation 11c (CD11c), but lower expression levels of CD80, CD86, and major histocompatibility complex class II. Nar-HSCs-imDCs also showed stronger inhibition of T cells and higher Treg cell proliferation. Interleukin 2 (IL-2) and interferon gamma levels were downregulated in Nar-HSCs-imDCs, whereas IL-4, IL-10, and transforming growth factor beta levels were upregulated. The rate of apoptosis and endocytic capacity of Nar-HSCs-DCs increased significantly after treatment with lipopolysaccharide. HSCs-imDCs or Nar-HSCs-imDCs were injected into Balb/c mice via the tail vein 7 days before skin grafting. Significantly reduced donor-specific CD4+ T cells and induced proliferation of CD4+CD25+FoxP3+ Treg cells were observed in the spleen of mice from the Nar-HSCs-imDCs group, especially at a dose of 106 Nar-HSCs-imDCs. The latter group also showed significantly prolonged survival of skin grafts.
Nar-HSCs-imDCs markedly improved the acceptance of organ allografts, offering a potentially new strategy for inducing immune tolerance in transplantation.
使用未成熟树突状细胞（imDCs）在刺激后诱导供体特异性免疫耐受受到其诱导率低和趋向成熟的限制。我们从骨髓造血干细胞（HSCs-imDCs）中获得 imDCs。然后，我们测试了柚皮素（Nar）抑制 HSCs-imDCs 成熟以诱导移植免疫耐受的能力。
从骨髓中收集 HSCs 并通过用 Nar 处理诱导其分化为 imDCs（Nar-HSCs-imDCs）。流式细胞术用于评估 DC 表面标志物、凋亡和内吞能力。混合淋巴细胞反应测定分析 DC 影响 T 细胞和调节性 T 细胞（Tregs）增殖的能力。酶联免疫吸附试验用于定量共培养的 DC 和 Tregs 上清液以及实验动物血清中的细胞因子水平。通过受体 Balb/c 小鼠的皮肤移植评估 Nar-HSCs-imDCs 诱导的免疫耐受水平，Kaplan-Meier 方法用于统计评估移植物存活。
与 HSC-imDCs 相比，Nar-HSCs-imDCs 表现出更高的分化簇 11c（CD11c）表达，但 CD80、CD86 和主要组织相容性复合体 II 水平较低。Nar-HSCs-imDCs 还表现出更强的 T 细胞抑制作用和更高的 Treg 细胞增殖。Nar-HSCs-imDCs 中白细胞介素 2（IL-2）和干扰素 γ 水平下调，而 IL-4、IL-10 和转化生长因子 β 水平上调。用脂多糖处理后，Nar-HSCs-DC 的凋亡率和内吞能力显著增加。HSCs-imDCs 或 Nar-HSCs-imDCs 在皮肤移植前 7 天通过尾静脉注入 Balb/c 小鼠。在 Nar-HSCs-imDCs 组的小鼠脾脏中观察到明显减少的供体特异性 CD4+T 细胞和诱导的 CD4+CD25+FoxP3+Treg 细胞增殖，特别是在 106 Nar-HSCs-imDCs 的剂量下。后者组还表现出明显延长的皮肤移植物存活时间。
Nar-HSCs-imDCs 显著改善了器官同种异体移植物的接受度，为诱导移植免疫耐受提供了一种新的潜在策略。
