Institute of Molecular Immunology, German Research Center for Environmental Health, Marchioninistrasse 25, 81377 Munich, Germany.
J Immunol. 2010 Jul 1;185(1):738-47. doi: 10.4049/jimmunol.1000060. Epub 2010 May 28.
In this paper, we describe a new method for preparation of human dendritic cells (DCs) that secrete bioactive IL-12(p70) using synthetic immunostimulatory compounds as TLR7/8 agonists. Monocyte-derived DCs were generated using a procedure that provided mature cells within 3 d. Several maturation mixtures that contained various cytokines, IFN-gamma, different TLR agonists, and PGE(2) were compared for impact on cell recovery, phenotype, cytokine secretion, migration, and lymphocyte activation. Mixtures that included the TLR7/8 agonists R848 or CL075, combined with the TLR3 agonist polyinosinic:polycytidylic acid, yielded 3-d mature DCs that secreted high levels of IL-12(p70), showed strong chemotaxis to CCR7 ligands, and had a positive costimulatory potential. They also had excellent capacity to activate NK cells, effectively polarized CD4(+) and CD8(+) T cells to secrete IFN-gamma and to induce T cell-mediated cytotoxic function. Thereby, mature DCs prepared within 3 d using such maturation mixtures displayed optimal functions required for vaccine development.
在本文中，我们描述了一种使用合成免疫刺激化合物作为 TLR7/8 激动剂制备分泌生物活性 IL-12(p70)的人树突状细胞 (DC) 的新方法。通过提供 3 天内成熟细胞的程序生成单核细胞来源的 DC。比较了几种包含不同细胞因子、IFN-γ、不同 TLR 激动剂和 PGE(2)的成熟混合物对细胞回收、表型、细胞因子分泌、迁移和淋巴细胞激活的影响。包含 TLR7/8 激动剂 R848 或 CL075 的混合物，与 TLR3 激动剂聚肌苷酸:聚胞苷酸相结合，产生 3 天成熟的 DC，分泌高水平的 IL-12(p70)，表现出对 CCR7 配体的强烈趋化性，并且具有正共刺激潜能。它们还具有有效激活 NK 细胞的能力，有效地极化 CD4(+)和 CD8(+)T 细胞分泌 IFN-γ，并诱导 T 细胞介导的细胞毒性功能。因此，使用这种成熟混合物在 3 天内制备的成熟 DC 显示出疫苗开发所需的最佳功能。
