Truxova Iva, Pokorna Katerina, Kloudova Kamila, Partlova Simona, Spisek Radek, Fucikova Jitka
Department of Immunology, Charles University, Second Faculty of Medicine and University Hospital Motol, Prague, Czech Republic; Sotio, Prague, Czech Republic.
Sotio, Prague, Czech Republic.
Immunol Lett. 2014 Jul;160(1):39-49. doi: 10.1016/j.imlet.2014.03.010. Epub 2014 Apr 12.
Dendritic cells (DCs) are professional antigen-presenting cells that are capable of inducing immune responses. DC-based vaccines are normally generated using a standard 5- to 7-day protocol. To shorten the DC-based vaccine production for use in cancer immunotherapy, we have developed a fast DC protocol by comparing standard DCs (Day 5 DCs) and fast DCs (Day 3 DCs).
We tested the generation of Day 5 versus Day 3 DCs using CellGro media and subsequent activation by two activation stimuli: Poly (I:C) and LPS. We evaluated DC morphology, viability, phagocyte activity, cytokine production and ability to stimulate antigen-specific T cells.
Day 5 and Day 3 DCs exhibited similar phagocytic capacity. Poly (I:C)-activated Day 5 DCs expressed higher levels of the costimulatory and surface molecules CD80, CD86 and HLA-DR compared to Poly (I:C)-activated Day 3 DCs. Nevertheless, LPS-activated Day 5 and Day 3 DCs were phenotypically similar. Cytokine production was generally stronger when LPS was used as the maturation stimulus, and there were no significant differences between Day 5 and Day 3 DCs. Importantly, Day 5 and Day 3 DCs were able to generate comparable numbers of antigen-specific CD8(+) T cells. The number of Tregs induced by Day 5 and Day 3 DCs was also comparable.
We identified monocyte-derived DCs generated in CellGro for 3 days and activated using Poly (I:C) similarly potent in most functional aspects as DCs produced by the standard 5 day protocol. These results provide the rationale for the evaluation of faster protocols for DC generation in clinical trials.
树突状细胞（DCs）是能够诱导免疫反应的专职抗原呈递细胞。基于DC的疫苗通常采用标准的5至7天方案制备。为了缩短用于癌症免疫治疗的基于DC的疫苗生产时间，我们通过比较标准DC（第5天DC）和快速DC（第3天DC）开发了一种快速DC方案。
我们使用CellGro培养基测试了第5天DC与第3天DC的生成情况，以及随后通过两种激活刺激物：聚肌胞苷酸（Poly (I:C)）和脂多糖（LPS）进行的激活。我们评估了DC的形态、活力、吞噬活性、细胞因子产生以及刺激抗原特异性T细胞的能力。
第5天和第3天的DC表现出相似的吞噬能力。与经聚肌胞苷酸激活的第3天DC相比，经聚肌胞苷酸激活的第5天DC表达更高水平的共刺激分子和表面分子CD80、CD86和HLA-DR。然而，经脂多糖激活的第5天和第3天DC在表型上相似。当使用脂多糖作为成熟刺激物时，细胞因子产生通常更强，并且第5天和第3天的DC之间没有显著差异。重要的是，第5天和第3天的DC能够产生相当数量的抗原特异性CD8(+) T细胞。第5天和第3天的DC诱导的调节性T细胞数量也相当。
我们确定了在CellGro中培养3天并使用聚肌胞苷酸激活的单核细胞衍生DC在大多数功能方面与标准5天方案产生的DC同样有效。这些结果为在临床试验中评估更快的DC生成方案提供了理论依据。
