Plasmid DNA uptake and subsequent cellular activation characteristics in human monocyte-derived cells in primary culture
Plasmid DNA uptake and subsequent cellular activation characteristics in human monocyte-derived cells in primary culture
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DOI：
10.1002/jps.20816
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发表时间：
2007-06-01
影响因子：
3.8
通讯作者：
Takakura, Yoshinobu
中科院分区：
文献类型：
--
作者：
Fukuhara, Yuga;Naoi, Tomoyuki;Takakura, Yoshinobu
在原代培养的人单核细胞、巨噬细胞和树突状细胞(DC)中，研究了三种类型的人单核细胞摄取质粒DNA(PDNA)和随后的细胞激活特性。裸露的pDNA与巨噬细胞和DC结合并被摄取，而只有单核细胞有明显的结合。多聚肌苷酸(Poly[I])、葡聚糖硫酸盐、马来酸化牛血清白蛋白(Mal-BSA)显著抑制PDNA与这些单核细胞来源的细胞的结合，多胞苷酸(Poly[C])对其抑制作用较小，但葡聚糖或半乳糖化BSA(Gal-BSA)、甘露糖化BSA(Man-BSA)对PDNA结合无明显影响，提示多聚阴离子参与了PDNA结合的一种特殊机制。在细胞激活研究中，无论PDNA中存在丰富的CpG基序，裸露的PDNA都不能诱导任何单核细胞产生肿瘤坏死因子Cz。然而，当与阳离子脂质体络合时，pDNA从人巨噬细胞产生大量的肿瘤坏死因子-α。单核细胞和DC均未观察到肿瘤坏死因子-α的诱导。此外，小牛胸腺DNA(CT DNA)也是络合的。使用阳离子脂质体也能诱导人巨噬细胞产生类似程度的肿瘤坏死因子-α。这些结果表明，在人单核细胞来源的细胞中，巨噬细胞被DNA以不依赖于CpG基序的方式与阳离子脂质体络合时被激活。(C)2007年Wiley-Liss，Inc.
Plasmid DNA (pDNA) uptake and subsequent cellular activation characteristics were studied in three types of human monocyte-derived cells, that is, human monocytes, macrophages, and dendritic cells (DCs) in primary culture. Naked pDNA was bound to and taken up by the macrophages and DCs while only significant binding occurred in the monocytes. pDNA binding to these monocyte-derived cells was significantly inhibited by polyinosinic acid (poly[I]), dextran sulfate, maleylated bovine serum albumin (Mal-BSA) and to a lesser extent by polycytidylic acid (poly[C]), but not by dextran or galactosylated BSA (Gal-BSA), mannosylated BSA (Man-BSA), suggesting that a specific mechanism for polyanions is involved in the pDNA binding. In cellular activation studies, naked pDNA could not induce TNF-cz production from any monocyte-derived cells, regardless of the abundant presence of CpG motifs in the pDNA. However, when complexed with cationic liposomes, pDNA produced a significant amount of TNF-alpha from the human macrophages. TNF-alpha induction was not observed in the monocytes or DCs. Moreover, calf thymus DNA (CT DNA) complexed. with cationic liposomes also induced TNF-alpha production to a similar extent in the human macrophages. These results indicate that, among human monocyte-derived cells, macrophages are activated by DNA when complexed with cationic liposomes in a CpG motif-independent manner. (C) 2007 Wiley-Liss, Inc.