Laboratory of Immunobiology for Research and Diagnosis (LIRAD), Blood and Tissue Bank (BST), Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Institut Investigació Germans Trias i Pujol (IGTP), Badalona, Barcelona, Spain.
Vaccine. 2012 Jan 5;30(2):378-87. doi: 10.1016/j.vaccine.2011.10.081. Epub 2011 Nov 12.
Immunotherapy using monocyte-derived dendritic cells (MDDC) is increasingly being considered as alternative therapeutic approach in cancer, infectious diseases and also in autoimmunity when patients are not responsive to conventional treatments. In general, generation of MDDC from monocytes is induced in the presence of GM-CSF and IL-4, and a maturation stimulus is added to the culture to obtain mature DCs suitable for therapy. For DC maturation, different combinations of pro-inflammatory mediators and Toll-like receptor ligands have been tested, obtaining DCs that differ in their properties and the type of immune response they promote. Therefore, it is necessary to find an optimal cytokine environment for DC maturation to obtain a cellular product suitable for DC-based immunotherapeutic protocols. In this study, we have evaluated in vitro the effects of different maturation stimuli on the viability, phenotype, cytokine profile, stability and functionality of immunogenic and tolerogenic (1α,25-dihydroxyvitamin D(3)-treated) MDDC. Maturation was induced using the clinical grade TLR4-agonist: monophosphoryl lipid A (LA), compared to the traditional cytokine cocktail (CC; clinical grade TNF-α, IL-1β, PGE2) and a combination of both. Our results showed the combination of CC+LA rendered a potent immunogenic DC population that induced the production of IFN-γ and IL-17 in allogeneic co-cultures, suggesting a Th17 polarization. Moreover, these immunogenic DCs showed a high surface expression of CD83, CD86, HLA-DR and secretion of IL-12p70. When aiming to induce tolerance, using LA to generate mature TolDC did not represent a clear advantage, and the stability and the suppressive capability exhibited by CC-matured TolDC may represent the best option. Altogether, these findings demonstrate the relevance of an appropriate maturation stimulus to rationally modulate the therapeutic potential of DCs in immunotherapy.
利用单核细胞衍生的树突状细胞（MDDC）进行免疫治疗，在癌症、传染病以及自身免疫性疾病中，当患者对常规治疗无反应时，作为一种替代治疗方法，其应用日益受到关注。通常，在 GM-CSF 和 IL-4 的存在下，从单核细胞中生成 MDDC，并向培养物中添加成熟刺激物，以获得适合治疗的成熟 DC。为了促进 DC 的成熟，已经测试了不同的促炎介质和 Toll 样受体配体的组合，获得了在其特性和促进的免疫反应类型上存在差异的 DC。因此，有必要为 DC 的成熟寻找最佳的细胞因子环境，以获得适合基于 DC 的免疫治疗方案的细胞产物。在这项研究中，我们评估了不同成熟刺激物对免疫原性和耐受性（1α，25-二羟维生素 D（3）处理）MDDC 的活力、表型、细胞因子谱、稳定性和功能的体外影响。成熟是通过临床级 TLR4 激动剂：单磷酰脂质 A（LA）诱导的，与传统细胞因子鸡尾酒（CC；临床级 TNF-α、IL-1β、PGE2）和两者的组合进行比较。我们的结果表明，CC+LA 的组合产生了一种有效的免疫原性 DC 群体，在同种异体共培养物中诱导 IFN-γ和 IL-17 的产生，提示 Th17 极化。此外，这些免疫原性 DC 表现出高表面表达 CD83、CD86、HLA-DR 和 IL-12p70 的分泌。当旨在诱导耐受性时，使用 LA 生成成熟的 TolDC 并没有明显的优势，而 CC 成熟的 TolDC 表现出的稳定性和抑制能力可能是最佳选择。总之，这些发现证明了适当的成熟刺激物对于合理调节 DC 在免疫治疗中的治疗潜力具有重要意义。
