Zhang Hui, Wang Yu, Wang Qian-Ting, Sun Sheng-Nan, Li Shi-You, Shang Hong, He You-Wen
NHC Key Laboratory of AIDS Immunology (China Medical University), Department of Laboratory Medicine, The First Affiliated Hospital of China Medical University, Shenyang, China.
National Clinical Research Center for Laboratory Medicine, The First Affiliated Hospital of China Medical University, Shenyang, China.
Front Cell Dev Biol. 2020 Mar 27;8:205. doi: 10.3389/fcell.2020.00205. eCollection 2020.
Dendritic cell (DC)-based vaccination is a promising immunotherapeutic strategy for cancer. However, clinical trials have shown only limited efficacy, suggesting the need to optimize protocols for human DC vaccine preparation. In this study, we systemically compared five different human DC vaccine maturation protocols used in clinical trials: (1) a four-cytokine cocktail (TNF-α, IL-6, IL-1β, and PGE2); (2) an α-DC-cytokine cocktail (TNF-α, IL-1β, IFN-α, IFN-γ, and poly I:C); (3) lipopolysaccharide (LPS)/IFN-γ; (4) TNF-α and PGE2; and (5) TriMix (mRNAs encoding CD40L, CD70, and constitutively active Toll-like receptor 4 electroporated into immature DCs). We found that the four-cytokine cocktail induced high levels of costimulatory and HLA molecules, as well as CCR7, in DCs. Mature DCs (mDCs) matured with the four-cytokine cocktail had higher viability than those obtained with the other protocols. Based on these features, we chose the four-cytokine cocktail protocol to further improve the immunizing capability of DCs by introducing exogenous genes. We showed that introducing exogenous Bcl-2 increased DC survival. Furthermore, introducing IL-12p70 rescued the inhibition of IL-12 secretion by PGE2 without impairing the DC phenotype. Introducing both Bcl-2 and IL-12p70 mRNAs into DCs induced enhanced cytomegalovirus pp65-specific CD8 T cells secreting IFN-γ and TNF-α. Taken together, our data suggest that DC matured by the four-cytokine cocktail combined with exogenous Bcl-2 and IL-12p70 gene expression represents a promising approach for clinical applications in cancer immunotherapy.
基于树突状细胞（DC）的疫苗接种是一种很有前景的癌症免疫治疗策略。然而，临床试验显示其疗效有限，这表明需要优化人DC疫苗制备方案。在本研究中，我们系统地比较了临床试验中使用的五种不同的人DC疫苗成熟方案：（1）四种细胞因子组合（TNF-α、IL-6、IL-1β和PGE2）；（2）α-DC细胞因子组合（TNF-α、IL-1β、IFN-α、IFN-γ和聚肌胞苷酸）；（3）脂多糖（LPS）/IFN-γ；（4）TNF-α和PGE2；以及（5）TriMix（编码CD40L、CD70和组成型活性Toll样受体4的mRNA电穿孔导入未成熟DC）。我们发现，四种细胞因子组合能诱导DC中高水平的共刺激分子和HLA分子以及CCR7。用四种细胞因子组合成熟的成熟DC（mDC）比用其他方案获得的mDC具有更高的活力。基于这些特性，我们选择四种细胞因子组合方案，通过引入外源基因进一步提高DC的免疫能力。我们发现，引入外源Bcl-2可提高DC的存活率。此外，引入IL-12p70可挽救PGE2对IL-12分泌的抑制作用，而不损害DC表型。将Bcl-2和IL-12p70 mRNA同时导入DC可诱导增强的分泌IFN-γ和TNF-α的巨细胞病毒pp65特异性CD8 T细胞。综上所述，我们的数据表明，通过四种细胞因子组合联合外源Bcl-2和IL-12p70基因表达而成熟的DC代表了一种在癌症免疫治疗中具有临床应用前景的方法。
