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

Pappalardo JS, Langellotti CA, Di Giacomo S, Olivera V, Quattrocchi V, Zamorano PI, Hartner WC, Levchenko TS, Torchilin VP - Int J Nanomedicine (2014)

Bottom Line: 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.The de novo synthesized gD protein was immunologically stimulating when transfections were performed with TATp-L, as indicated by the secretion of interleukin 6.

View Article: PubMed Central - PubMed

Affiliation: Virology Institute, Center for Research in Veterinary and Agronomic Sciences, National Institute for Agricultural Technology (INTA), Hurlingham, BA, Argentina ; National Council for Scientific and Technical Research (CONICET), Autonomous City of Buenos Aires, Argentina ; Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, Boston, MA, USA.

ABSTRACT
Dendritic cells (DC) are antigen-presenting cells uniquely capable of priming naïve T cells and cross-presenting antigens, and they 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 a higher expression of the enhanced green fluorescent protein and bovine herpes virus type 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.

Show MeSH

Related in: MedlinePlus

Evaluation of GM-CSF-stimulated bone marrow-derived cells cultured (loosely attached cells) for the expression of CD11c.Note: Typically 65% or more of the cells were positive for this DC marker (solid histogram, FITC-CD11c) in comparison with the isotype control (transparent histogram).Abbreviations: CD11c, cluster of differentiation 11c; DC, dendritic cells; FITC, fluorescein isothiocyanate; GM-CSF, granulocyte-macrophage colony-stimulating factor.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3928453&req=5

f1-ijn-9-963: Evaluation of GM-CSF-stimulated bone marrow-derived cells cultured (loosely attached cells) for the expression of CD11c.Note: Typically 65% or more of the cells were positive for this DC marker (solid histogram, FITC-CD11c) in comparison with the isotype control (transparent histogram).Abbreviations: CD11c, cluster of differentiation 11c; DC, dendritic cells; FITC, fluorescein isothiocyanate; GM-CSF, granulocyte-macrophage colony-stimulating factor.

Mentions: Flow cytometry of the bone marrow-derived DC cultures demonstrated that more than 60% of the loosely adherent cells exhibited specific molecular markers for DC (CD11c), which provided a sufficient level of cell purity for these studies (Figure 1). MHC class II marker was present (20%) and Gr-1-positive cells constituted less than 10% of all cells (data not shown).


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

Pappalardo JS, Langellotti CA, Di Giacomo S, Olivera V, Quattrocchi V, Zamorano PI, Hartner WC, Levchenko TS, Torchilin VP - Int J Nanomedicine (2014)

Evaluation of GM-CSF-stimulated bone marrow-derived cells cultured (loosely attached cells) for the expression of CD11c.Note: Typically 65% or more of the cells were positive for this DC marker (solid histogram, FITC-CD11c) in comparison with the isotype control (transparent histogram).Abbreviations: CD11c, cluster of differentiation 11c; DC, dendritic cells; FITC, fluorescein isothiocyanate; GM-CSF, granulocyte-macrophage colony-stimulating factor.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928453&req=5

f1-ijn-9-963: Evaluation of GM-CSF-stimulated bone marrow-derived cells cultured (loosely attached cells) for the expression of CD11c.Note: Typically 65% or more of the cells were positive for this DC marker (solid histogram, FITC-CD11c) in comparison with the isotype control (transparent histogram).Abbreviations: CD11c, cluster of differentiation 11c; DC, dendritic cells; FITC, fluorescein isothiocyanate; GM-CSF, granulocyte-macrophage colony-stimulating factor.
Mentions: Flow cytometry of the bone marrow-derived DC cultures demonstrated that more than 60% of the loosely adherent cells exhibited specific molecular markers for DC (CD11c), which provided a sufficient level of cell purity for these studies (Figure 1). MHC class II marker was present (20%) and Gr-1-positive cells constituted less than 10% of all cells (data not shown).

Bottom Line: 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.The de novo synthesized gD protein was immunologically stimulating when transfections were performed with TATp-L, as indicated by the secretion of interleukin 6.

View Article: PubMed Central - PubMed

Affiliation: Virology Institute, Center for Research in Veterinary and Agronomic Sciences, National Institute for Agricultural Technology (INTA), Hurlingham, BA, Argentina ; National Council for Scientific and Technical Research (CONICET), Autonomous City of Buenos Aires, Argentina ; Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, Boston, MA, USA.

ABSTRACT
Dendritic cells (DC) are antigen-presenting cells uniquely capable of priming naïve T cells and cross-presenting antigens, and they 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 a higher expression of the enhanced green fluorescent protein and bovine herpes virus type 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.

Show MeSH
Related in: MedlinePlus