Plasmid DNA Uptake and Subsequent Cellular Activation Characteristics in Human Monocyte-Derived Cells in Primary Culture

• Published in: Journal of pharmaceutical sciences 2007-06, Vol.96 (6), p.1576-1584
• Main Authors: Fukuhara, Yuga, Naoi, Tomoyuki, Ogawa, Yoshiyuki, Nishikawa, Makiya, Takakura, Yoshinobu
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Cells, Cultured; cellular uptake; CpG Islands; CT DNA, calf thymusDNA; DC, dendritic cell; degradation; DNA - metabolism; gene therapy; General pharmacology; Humans; Medical sciences; monocyte-derived cells; Monocytes - metabolism; pDNA, plasmid DNA; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; plasmid DNA; Plasmids; poly[C], polycytidylic acidxy; poly[I], polyinosinic acid; Polymers - pharmacology; TLR, toll-like receptor; Tumor Necrosis Factor-alpha - biosynthesis
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1002/jps.20816

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‐α 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‐α from the human macrophages. TNF‐α induction was not observed in the monocytes or DCs. Moreover, calf thymus DNA (CT DNA) complexed with cationic liposomes also induced TNF‐α 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.