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Polyethyleneimine‐based immunopolyplex for targeted gene transfer in human lymphoma celllines
Author(s) -
Guillem Vicent M.,
Tormo Mar,
Revert Fernando,
Benet Isabel,
GarcíaConde Javier,
Crespo Antonio,
Aliño Salvador F.
Publication year - 2002
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.228
Subject(s) - transfection , jurkat cells , cd19 , gene delivery , microbiology and biotechnology , cd3 , reporter gene , cell culture , biology , flow cytometry , t cell , chemistry , virology , gene , antigen , biochemistry , gene expression , immune system , immunology , cd8 , genetics
Background Specific and efficient delivery of genes into targeted cells is a priority objective in non‐viral gene therapy. Polyethyleneimine‐based polyplexes have been reported to be good non‐viral transfection reagents. However, polyplex‐mediated DNA delivery occurs through a non‐specific mechanism. This article reports the construction of an immunopolyplex, a targeted non‐viral vector based on a polyplex backbone, and its application in gene transfer over human lymphoma cell lines. Methods Targeting elements (biotin‐labeled antibodies), which should recognize a specific element of the target cell membrane and promote nucleic acid entry into the cell, were attached to the polyplex backbone through a bridge protein (streptavidin). Immunopolyplex transfection activity was studied in several hematological cell lines [Jurkat (CD3+/CD19−), Granta 519 (CD3−/ CD19+), and J.RT3‐T3.5 (CD3−/CD19−)] using the EGFP gene as a reporter gene and anti‐CD3 and anti‐CD19 antibodies as targeting elements. Transfection activity was evaluated via green fluorescence per cell and the percentage of positive cells determined by flow cytometry. Results A significant selectivity of gene delivery was observed, since the anti‐CD3 immunopolyplex worked only in Jurkat cells while the anti‐CD19 immunopolyplex worked only in the Granta cell line. Moreover, transfection of a CD3+/CD3− cell mixture with anti‐CD3 immunopolyplexes showed up to 16‐fold more transfection in CD3+ than in CD3− cells. Several non‐specific transfection reagents showed poor or no transfection activity. Conclusion It is concluded that immunopolyplex is a good non‐viral vector for specific and selective nucleic acid delivery. Immunopolyplex design allows easy replacement of the targeting element (antibody) – the streptavidin–polyplex backbone remaining intact – thereby conferring high versatility. Copyright © 2002 John Wiley & Sons, Ltd.