Improved transfection of spleen-derived antigen-presenting cells in culture using TATp-liposomes.
Antigen presenting cells (APC) are among the most important cells of the immune system since they link the innate and the adaptative immune responses, directing the type of immune response to be elicited. To modulate the immune response in immune preventing or treating therapies, gene delivery into immunocompetent cells could be used. However, APC are very resistant to transfection. To increase the efficiency of APC transfection, we have used liposome-based lipoplexes additionally modified with cell-penetrating TAT peptide (TATp) for better intracellular delivery of a model plasmid encoding for the enhanced-green fluorescent protein (pEGFP). pEGFP-bearing lipoplexes made of a mixture of PC:Chol:DOTAP (60:30:10 molar ratio) with the addition of 2% mol of polyethylene glycol-phosphatidylethanolamine (PEG-PE) conjugate (plain-L) or TATp-PEG-PE (TATp-L) were shown to effectively protect the incorporated DNA from degradation. Uptake assays of rhodamine-labeled lipoplexes and transfections with the EGFP reporter gene were performed with APC derived from the mouse spleen. TATp-L-based lipoplexes allowed for significantly enhanced both, the uptake and transfection in APC. Such a tool could be used for the APC transfection as a first step in immune therapy.
Pappalardo
Juan Sebastián
JS
Instituto de Virología, Centro de Investigaciones en Ciencias Veterinarias y Agronómicas, INTA-Castelar, Buenos Aires, Argentina.
Quattrocchi
Valeria
V
Langellotti
Cecilia
C
Di Giacomo
Sebastián
S
Gnazzo
Victoria
V
Olivera
Valeria
V
Calamante
Gabriela
G
Zamorano
Patricia I
PI
Levchenko
Tatyana S
TS
Torchilin
Vladimir P
VP
D000818:Animals; D000938:Antigen-Presenting Cells / Q000166:cytology* / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry; D015696:Gene Products, tat / Q000737:chemistry / Q000235:genetics; D008081:Liposomes / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D015696:Gene Products, tat; D008081:Liposomes; D004247:DNA