Division of Cardiovascular and Rare Disease, Korea Center for Disease Control and Prevention 194 Tongillo, Eunpyung-gu, Seoul, Republic of Korea.
Vaccine. 2010 Apr 1;28(16):2877-86. doi: 10.1016/j.vaccine.2010.01.061. Epub 2010 Feb 13.
Effective induction of cell-mediated immune responses strongly depends on the ability of dendritic cells (DCs) to produce Th1-polarizing cytokines, migrate to lymph nodes and stimulate T cells through antigen-presenting complex and costimulatory molecules. While various protocols for optimizing DC maturation with single or multiple stimuli mimicking infections or inflammatory milieu have been proposed for the generation of DCs with features desired for clinical application, stepwise maturation of DCs by these multiple stimuli has not been systemically assessed. Among the combinations of several immune-modulating factors with known effects on DC maturation, we found that stepwise DC maturation with cytokine cocktail (TNF-alpha+IL-6+IL-1 beta+PGE(2)) followed by poly(I:C) stimulation enhanced the production of IL-12 with strong allostimulatory capacity. While there were no significant differences between DC matured by simultaneous or sequential activation by cytokine cocktail and poly(I:C) in expression of markers and costimulatory molecules of mature DCs, the delivery of inflammatory signal prior to poly(I:C) results in sustained interleukin-12 expression with reduced IL-10 than DC matured by simultaneous stimulation. This sequential stimulation significantly increased migratory capacity in response to CCL21 and CXCL12 compared to DC matured with cytokine cocktail. Furthermore, these DCs retained their responsiveness to CD40L stimulation in secondary IL-12 production and efficiently generated autologous antigen-specific effector T cells as evidenced by ELISPOT assay. Thus, we propose a novel DC maturation protocol in which stimulation of DCs with cytokine cocktail and subsequently with poly(I:C) generates DCs with a high migratory capacity with a preferential Th1 inducing capacity.
有效的细胞介导免疫反应的诱导强烈依赖于树突状细胞 (DCs) 产生 Th1 极化细胞因子的能力、迁移到淋巴结并通过抗原呈递复合物和共刺激分子刺激 T 细胞的能力。虽然已经提出了各种优化 DC 成熟的方案，使用模拟感染或炎症环境的单一或多种刺激物来产生具有临床应用所需特征的 DC，但这些多种刺激物对 DC 成熟的逐步作用尚未得到系统评估。在几种已知对 DC 成熟有影响的免疫调节因子的组合中，我们发现用细胞因子鸡尾酒 (TNF-α+IL-6+IL-1β+PGE(2)) 逐步成熟 DC，然后用 poly(I:C) 刺激可增强具有强烈同种刺激能力的 IL-12 的产生。虽然细胞因子鸡尾酒和 poly(I:C) 同时或顺序激活成熟的 DC 之间在成熟 DC 的标志物和共刺激分子表达方面没有显著差异，但在 poly(I:C) 之前传递炎症信号会导致 IL-12 的持续表达，IL-10 减少比同时刺激成熟的 DC。与用细胞因子鸡尾酒成熟的 DC 相比，这种顺序刺激显著增加了对 CCL21 和 CXCL12 的迁移能力。此外，这些 DC 在二次 IL-12 产生中保持对 CD40L 刺激的反应性，并通过 ELISPOT 测定有效地产生了自体抗原特异性效应 T 细胞。因此，我们提出了一种新的 DC 成熟方案，即用细胞因子鸡尾酒刺激 DC，随后用 poly(I:C) 刺激，可产生具有高迁移能力和优先诱导 Th1 能力的 DC。
