title: Polyethyleneimine-based immunopolyplex for targeted gene transfer in human lymphoma cell lines.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Servei d'Hematologia i Oncologia, Hospital Clínic Universitari, Facultat de Medicina, Universitat de València, Avda. Blasco Ibáñez 17, 46010 València, Spain.
authors.forename: Vicent M
authors.initials: VM
authors.lastname: Guillem
authors.forename: Mar
authors.initials: M
authors.lastname: Tormo
authors.forename: Fernando
authors.initials: F
authors.lastname: Revert
authors.forename: Isabel
authors.initials: I
authors.lastname: Benet
authors.forename: Javier
authors.initials: J
authors.lastname: García-Conde
authors.forename: Antonio
authors.initials: A
authors.lastname: Crespo
authors.forename: Salvador F
authors.initials: SF
authors.lastname: Aliño
mesh_terms: D018941:Antigens, CD19 / Q000096:biosynthesis; D019921:Biotinylation; D017252:CD3 Complex / Q000096:biosynthesis; D004247:DNA / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D019169:Jurkat Cells; D008164:Luminescent Proteins / Q000378:metabolism; D008223:Lymphoma / Q000276:immunology* / Q000628:therapy; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011094:Polyethyleneimine / Q000737:chemistry*; D011485:Protein Binding; D019809:Streptavidin / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D018941:Antigens, CD19; D017252:CD3 Complex; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D004247:DNA; D019809:Streptavidin; D003851:Deoxyribonucleases