In vitro transfection of bone marrow-derived dendritic cells with TATp-liposomes

Abstract

Dendritic cells (DC) are antigen presenting cells (APC) uniquely capable of priming naïve T cells and cross-presenting antigens and determine the type of immune response elicited against an antigen. TAT peptide (TATp), is an amphipathic, arginine-rich, cationic peptide that promotes penetration and translocation of various molecules and nanoparticles into cells. TATp-liposomes (TATp-L) used for DC transfection were prepared using TATp derivatized with a lipid-terminated polymer capable of anchoring in the liposomal membrane. Here, we show that the addition of TATp to DNA-loaded liposomes increased the uptake of DNA in DC. DNA-loaded TATp-L increased the in vitro transfection efficiency in DC cultures as evidenced by an enhanced expression of the enhanced green fluorescent protein (EGFP) and bovine herpes virus type 1 (BoHV-1) glycoprotein D (gD). The de novo synthesized gD protein was immunologically stimulating when transfections were performed with TATp-L as indicated by the secretion of interleukin 6 (IL-6).