通过多聚泛素化靶向蛋白酶体的mRNA树突状细胞疫苗。

Dendritic cell vaccine with mRNA targeted to the proteasome by polyubiquitination.

作者信息

Hosoi Akihiro, Takeda Yayoi, Sakuta Kazuko, Ueha Satoshi, Kurachi Makoto, Kimura Kiminori, Maekawa Ryuji, Kakimi Kazuhiro

机构信息

Department of Immunotherapeutics (Medinet), Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-Ku, Tokyo 113-8655, Japan.

出版信息

Biochem Biophys Res Commun. 2008 Jun 27;371(2):242-6. doi: 10.1016/j.bbrc.2008.04.034. Epub 2008 Apr 28.

DOI:10.1016/j.bbrc.2008.04.034

PMID:18423376

Abstract

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 20 ng/ml GM-CSF and maturation with 1 microg/ml LPS. These mDCs were then electroporated with 10 microg 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.

摘要

用编码肿瘤相关抗原（TAA）的mRNA转染的树突状细胞（DC）可诱导肿瘤特异性T细胞反应。为增强这一效果，我们用靶向蛋白酶体的TAA编码mRNA转染成熟DC（mDC）。通过用20 ng/ml粒细胞-巨噬细胞集落刺激因子（GM-CSF）培养从骨髓细胞生成DC，并以1 μg/ml脂多糖（LPS）使其成熟。然后用10 μg mRNA对这些mDC进行电穿孔。将体外转录mRNA电穿孔后的抗原呈递与泛素前导的TAA构建体的mRNA进行比较。发现编码共翻译泛素化抗原的mRNA的蛋白酶体靶向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，我们认为通过使用靶向蛋白酶体的mRNA可提高RNA转染的DC疫苗效力。