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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.

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Related in: MedlinePlus

Dendritic cells after 10-minutes incubation with TATp-L and 150 minutes in normal medium.Notes: Two serial photographs were focused in parallel tangential (A) and longitudinal (B) planes. The vesicle-like compartments were observed at 100× with digital augmentation. Reference marker =10 μm.Abbreviation: TATp-L, TAT peptide liposomes.
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f4-ijn-9-963: Dendritic cells after 10-minutes incubation with TATp-L and 150 minutes in normal medium.Notes: Two serial photographs were focused in parallel tangential (A) and longitudinal (B) planes. The vesicle-like compartments were observed at 100× with digital augmentation. Reference marker =10 μm.Abbreviation: TATp-L, TAT peptide liposomes.

Mentions: To demonstrate that TATp-L penetrated DC, serial pictures of the treated cells that showed rhodamine fluorescence were taken in two parallel planes (tangential and longitudinal) after 150 minutes of incubation in complete medium. The images clearly demonstrated vesicles with Rh-labeled TATp-L inside cells (Figure 4), which confirmed the enhanced penetration of DC by TATp-L.


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)

Dendritic cells after 10-minutes incubation with TATp-L and 150 minutes in normal medium.Notes: Two serial photographs were focused in parallel tangential (A) and longitudinal (B) planes. The vesicle-like compartments were observed at 100× with digital augmentation. Reference marker =10 μm.Abbreviation: TATp-L, TAT peptide liposomes.
© Copyright Policy
Related In: Results  -  Collection

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

f4-ijn-9-963: Dendritic cells after 10-minutes incubation with TATp-L and 150 minutes in normal medium.Notes: Two serial photographs were focused in parallel tangential (A) and longitudinal (B) planes. The vesicle-like compartments were observed at 100× with digital augmentation. Reference marker =10 μm.Abbreviation: TATp-L, TAT peptide liposomes.
Mentions: To demonstrate that TATp-L penetrated DC, serial pictures of the treated cells that showed rhodamine fluorescence were taken in two parallel planes (tangential and longitudinal) after 150 minutes of incubation in complete medium. The images clearly demonstrated vesicles with Rh-labeled TATp-L inside cells (Figure 4), which confirmed the enhanced penetration of DC by TATp-L.

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