Stable nonviral gene transfer into primary human T cells.
Effective techniques for the stable genetic modification of peripheral T cells would facilitate functional gene studies and the development of gene therapeutic approaches. However, many approaches to genetically modify T cells are hampered by low transfection efficiency, direct cell toxicity, and the need for specialized laboratory space. In this study we investigated the Amaxa Nucleofector platform, a nonviral technique to transfect primary human T cells. A plasmid equipped with two different promoters enabled concomitant expression of a gene of interest and of a cell surface marker allowing for immunomagnetic cell enrichment. This resulted in highly purified populations of gene-modified T cells and, after repeated enrichment steps, provided stably and homogeneously transfected, fully functional human T cells. In summary, this study provides proof of principle that human T cells can be altered to homogeneously and stably express a gene of interest by a nonviral technique. This should enable further studies on T cell physiology and ultimately facilitate the translation of treatment approaches either for diseases that are caused by defective gene function in T cells or for diseases that require genetically designed T cell therapy.
Magg
T
T
Department of Pediatric Hematology/Oncology, Dr. von Haunersches Kinderspital, Munich, Germany.
Hartrampf
S
S
Albert
M H
MH
D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation; D008213:Lymphocyte Activation; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D009414:Nerve Growth Factors; D049452:Green Fluorescent Proteins