Development of a DNA Vaccine for Melanoma Metastasis by Inhalation Based on an Analysis of Transgene Expression Characteristics of Naked pDNA and a Ternary Complex in Mouse Lung Tissues
Development of a DNA Vaccine for Melanoma Metastasis by Inhalation Based on an Analysis of Transgene Expression Characteristics of Naked pDNA and a Ternary Complex in Mouse Lung Tissues
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DOI：
10.3390/pharmaceutics12060540
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发表时间：
2020-06-01
期刊：
影响因子：
5.4
通讯作者：
Nakamura, Tadahiro
中科院分区：
文献类型：
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作者：
Kodama, Yukinobu;Nakashima, Mikiro;Nakamura, Tadahiro
本研究探讨了质粒DNA（pDNA）肺部给药系统及其在黑色素瘤DNA疫苗中的应用。使用pCMV-Luc、pEGFP-C1和pZsGreen作为模型pDNA，以评估小鼠吸入后的转染功效。裸pDNA和三元复合物，由pDNA，树枝状移植物聚-L-赖氨酸（DGL），和γ-聚谷氨酸（γ-PGA），都表现出强烈的基因表达后吸入肺。通过使用多色深度成像观察，检测肺泡巨噬细胞富集部位的转基因表达。在这些结果的基础上，我们使用表达黑色素瘤相关抗原（泛素化鼠黑色素瘤gp 100和酪氨酸酶相关蛋白2（TRP 2）肽表位）的pUb-M作为黑色素瘤的DNA疫苗。吸入裸pUb-M及其三元复合物可显著抑制小鼠黑色素瘤细胞系B16-F10的转移。增强Th 1应答的炎性细胞因子如TNF-α、IFN-γ和IL-6的水平在pUb-M三元复合物中高于裸pUb-M和pEGFP-C1三元复合物作为对照。总之，我们澄清了吸入裸pDNA及其三元复合物是一种有用的癌症疫苗接种技术。
The present study investigated a pulmonary delivery system of plasmid DNA (pDNA) and its application to melanoma DNA vaccines. pCMV-Luc, pEGFP-C1, and pZsGreen were used as a model pDNA to evaluate transfection efficacy after inhalation in mice. Naked pDNA and a ternary complex, consisting of pDNA, dendrigraft poly-l-lysine (DGL), and gamma-polyglutamic acid (gamma-PGA), both showed strong gene expression in the lungs after inhalation. The transgene expression was detected in alveolar macrophage-rich sites by observation using multi-color deep imaging. On the basis of these results, we used pUb-M, which expresses melanoma-related antigens (ubiquitinated murine melanoma gp100 and tyrosinase-related protein 2 (TRP2) peptide epitopes), as DNA vaccine for melanoma. The inhalation of naked pUb-M and its ternary complex significantly inhibited the metastasis of B16-F10 cells, a melanoma cell line, in mice. The levels of the inflammatory cytokines, such as TNF-alpha, IFN-gamma, and IL-6, which enhance Th1 responses, were higher with the pUb-M ternary complex than with naked pUb-M and pEGFP-C1 ternary complex as control. In conclusion, we clarified that the inhalation of naked pDNA as well as its ternary complex are a useful technique for cancer vaccination.