Enhancing the T-cell stimulatory capacity of human dendritic cells by co-electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA
Enhancing the T-cell stimulatory capacity of human dendritic cells by co-electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA
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DOI：
10.1038/mt.2008.77
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发表时间：
2008-06-01
影响因子：
12.4
通讯作者：
Thielemans, Kris
中科院分区：
文献类型：
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作者：
Bonehill, Aude;Tuyaerts, Sandra;Thielemans, Kris
目前使用的树突状细胞（DC）疫苗接种方案的有效性可以通过为DC提供更有效的成熟信号来提高。因此，我们研究了人单核细胞来源的DC的T细胞刺激能力是否可以通过与CD 40 L、CD 70和组成型活性Toll样受体4（caTLR 4）编码mRNA的不同组合共电穿孔来增加。我们表明，未成熟的DC电穿孔与CD 40 L和/或CaTLR 4 mRNA，但不是那些电穿孔与CD 70 mRNA，获得一个成熟的表型沿着增强分泌的几种细胞因子/趋化因子。此外，这些DC在诱导幼稚CD 4（+）T细胞分化为分泌干扰素-γ（IFN-γ）的1型T辅助（Th 1）细胞方面非常有效。此外，我们评估了电穿孔的DC在不添加任何外源性细胞因子的情况下激活幼稚HLA-A2限制性MelanA特异性CD 8（+）T细胞的能力。当所有三种分子组合时，与未成熟DC相比，观察到MelanA特异性CD 8（+）T细胞增加>500倍，与细胞因子混合物成熟DC相比，增加>200倍。作为相关性，我们发现溶细胞性和分泌IFN-γ/肿瘤坏死因子-α（TNF-alpha）的CD 8（+）T细胞显着增加。我们的数据表明，基因修饰表达刺激分子的未成熟DC可以在体外诱导肿瘤抗原特异性T细胞，并且可以证明与目前使用的方法成熟的DC相比有显着改善。
The effectiveness of the dendritic cell (DC) vaccination protocols that are currently in use could be improved by providing the DCs with a more potent maturation signal. We therefore investigated whether the T-cell stimulatory capacity of human monocyte-derived DCs could be increased by co-electroporation with different combinations of CD40L, CD70, and constitutively active toll-like receptor 4 (caTLR4) encoding mRNA. We show that immature DCs electroporated with CD40L and/or caTLR4 mRNA, but not those electroporated with CD70 mRNA, acquire a mature phenotype along with an enhanced secretion of several cytokines/chemokines. Moreover, these DCs are very potent in inducing naive CD4(+) T cells to differentiate into interferon-gamma (IFN-gamma)-secreting type 1 T helper (Th1) cells. Further, we assessed the capacity of the electroporated DCs to activate naive HLA-A2-restricted MelanA-specific CD8(+) T cells without the addition of any exogenous cytokines. When all three molecules were combined, a >500-fold increase in MelanA-specific CD8(+) T cells was observed when compared with immature DCs, and a >200-fold increase when compared with cytokine cocktail-matured DCs. In correlation, we found a marked increase in cytolytic and IFN-gamma/tumor necrosis factor-alpha (TNF-alpha) secreting CD8(+) T cells. Our data indicate that immature DCs genetically modified to express stimulating molecules can induce tumor antigen-specific T cells in vitro and could prove to be a significant improvement over DCs matured with the methods currently in use.