Dendritic cell vaccine with mRNA targeted to the proteasome by polyubiquitination.
Dendritic cell vaccine with mRNA targeted to the proteasome by polyubiquitination.
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DOI：
10.1016/j.bbrc.2008.04.034
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发表时间：
2008-06
影响因子：
3.1
通讯作者：
A. Hosoi;Y. Takeda;K. Sakuta;S. Ueha;M. Kurachi;K. Kimura;R. Maekawa;K. Kakimi
中科院分区：
文献类型：
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作者：
A. Hosoi;Y. Takeda;K. Sakuta;S. Ueha;M. Kurachi;K. Kimura;R. Maekawa;K. Kakimi
转染编码肿瘤相关抗原 (TAA) 的 mRNA 的树突状细胞 (DC) 可以诱导肿瘤特异性 T 细胞反应。为了加强这一点，我们用编码靶向蛋白酶体的 TAA 的 mRNA 转染成熟 DC (mDC)。通过用 20ng/ml GM-CSF 培养并用 1μg/ml LPS 成熟，从骨髓细胞产生 DC。然后用 10μg mRNA 对这些 mDC 进行电穿孔。将体外转录的 mRNA 电穿孔后的抗原呈递与来自泛素之前的 TAA 构建体的 mRNA 进行比较。研究发现，蛋白酶体靶向编码共翻译泛素化抗原的 mRNA 可增强靶蛋白的细胞内降解，并导致 TAA 特异性 CD8+T 细胞更有效的启动和扩增。因此，我们建议通过使用靶向蛋白酶体的 mRNA 可以提高 RNA 转染的 DC 疫苗的功效。
Dendritic cells (DCs) transfected with mRNA encoding tumor-associated antigens (TAAs) can induce tumor-specific T-cell responses. To potentiate this, we transfected mature DCs (mDCs) with mRNA encoding TAA targeted to the proteasome. DCs were generated from bone marrow cells by culture with 20ng/ml GM-CSF and maturation with 1μg/ml LPS. These mDCs were then electroporated with 10μg of mRNA. Antigen presentation after electroporation with in vitro transcribed mRNA was compared with mRNA from a construct of the TAA preceded by ubiquitin. Proteasomal targeting of mRNA encoding cotranslationally ubiquitinated antigen was found to enhance intracellular degradation of target protein, and result in more efficient priming and expansion of TAA-specific CD8+T-cells. We therefore suggest that RNA-transfected DC vaccine efficacy could be improved by the use of mRNA targeted to the proteasome.