title: Gene Modification of Human Natural Killer Cells Using a Retroviral Vector.
abstract: As part of the innate immune system, natural killer (NK) cells are regarded as promising effector cells for adoptive cell therapy approaches to treat patients with cancer. In some cases, genetic modification of the NK cells may be considered but such manipulation has to be integrated into the expansion method to allow the generation of clinically relevant numbers of gene-modified NK cells. Therefore, an efficient gene transfer procedure is needed.Our group developed a retrovirus-based transduction protocol capable of robust expansion of gene-modified NK cells with a high rate of transgene expression. Actively dividing cells is a prerequisite for efficient gene transfer when using a retroviral vector. In the procedure presented here, strong activation of the NK cells was provided by a combination of IL-15 and the K-562 feeder cells. Beside the interest in developing a simple procedure compliant with good manufacturing practice (GMP) for the production of therapeutic products, this approach also provides a valuable means of generating genetically modified primary NK cells for future preclinical studies.
authors.affiliation: Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., P14.2920, Unit #065, Houston, TX, 77030, USA.
authors.forename: Joshua N
authors.initials: JN
authors.lastname: Kellner
authors.affiliation: Program for Cell Enhancement and Technologies for Immunotherapy, The Sheikh Zayed Institute for Pediatric Surgical Innovation, the Center for Cancer and Immunology Research, Children's National Health System, Washington, DC, 20052, USA.|Departments of Pediatrics and Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC, 20052, USA.
authors.forename: Conrad R
authors.initials: CR
authors.lastname: Cruz
authors.affiliation: Program for Cell Enhancement and Technologies for Immunotherapy, The Sheikh Zayed Institute for Pediatric Surgical Innovation, the Center for Cancer and Immunology Research, Children's National Health System, Washington, DC, 20052, USA.|Departments of Pediatrics and Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC, 20052, USA.
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Bollard
authors.affiliation: Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., P14.2920, Unit #065, Houston, TX, 77030, USA. ESYvon@mdanderson.org.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Yvon
mesh_terms: D002478:Cells, Cultured; D061252:Feeder Cells / Q000166:cytology / Q000276:immunology; D005822:Genetic Vectors; D006801:Humans; D019409:Interleukin-15 / Q000235:genetics* / Q000378:metabolism; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: C569274:IL15 protein, human; D019409:Interleukin-15
keywords: Gene transfer; K-562 feeder cells; NK cells expansion; Natural killer cells; Retroviral vector; Single transduction