A cationic peptide consists of ornithine and histidine repeats augments gene transfer in dendritic cells
A cationic peptide consists of ornithine and histidine repeats augments gene transfer in dendritic cells
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DOI：
10.1016/j.molimm.2003.08.001
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发表时间：
2003-12-01
影响因子：
3.6
通讯作者：
Meng, WS
中科院分区：
文献类型：
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作者：
Chamarthy, SP;Kovacs, JR;Meng, WS
将质粒与聚-L-赖氨酸（PLL）和聚-L-鸟氨酸（PLO）等高分子量阳离子聚合物缩合可以增强用编码抗原的裸露DNA进行基因疫苗接种所产生的抗原特异性免疫。虽然这些高分子量聚合物在转染实验中明显有效，但其物理异质性和毒性限制了临床应用。在树突状细胞 (DC) 中的 DNA 结合、毒性和基因转移效率的背景下，对三种化学定义的低分子量阳离子肽 K-16、K10H6 和 O10H6 进行了检查。结果表明，虽然所有三种肽都能以相似的效率结合编码报告基因的质粒，但用与 O10H6 复合的 DNA 进行体外转染可产生最高水平的基因表达。此外，游离的O10H6对DC没有毒性，而基于赖氨酸的肽在DC培养物中引起显着的细胞死亡。我们还表明，用与 O10H6 复合的 DNA 离体转染的 DC 能够在体内引发抗原特异性 INFgamma 产生。综上所述，这些结果表明鸟氨酸和组氨酸重复序列是 DC 非病毒基因转移载体的合适构建模块。 (C) 2003 Elsevier Ltd. 保留所有权利。
Condensing the plasmid with high molecular weight cationic polymers such as poly-L-lysine (PLL) and poly-L-ornithine (PLO) can enhance antigen-specific immunity generated from genetic vaccination with naked DNA encoding antigens. While these high molecular weight polymers are clearly effective in transfection experiments, clinical applications are limited by their physical heterogeneity and toxicity. Three chemically defined low molecular weight cationic peptides, K-16, K10H6, and O10H6, were examined in the context of DNA binding, toxicity, and efficiency of gene transfer in dendritic cells (DC). The results showed that while all three peptides can bind to a plasmid encoding a reporter gene with similar efficiency, in vitro transfection with DNA complexed with O10H6 complexed resulted in the highest level of gene expression. Moreover, free O10H6 was not toxic to DC, while the lysine-based peptides caused significant cell death in DC cultures. We also showed that DC transfected ex vivo with DNA complexed with O10H6 was capable of eliciting antigen-specific INFgamma production in vivo. Taken together, these results indicate ornithine and histidine repeats are suitable building blocks of non-viral gene transfer vector for DC. (C) 2003 Elsevier Ltd. All rights reserved.