title: Genetic correction of cultured T cells from an adenosine deaminase-deficient patient: characteristics of non-transduced and transduced T cells.
abstract: T lymphocytes derived from peripheral blood of a patient with adenosine deaminase (ADA) deficiency were expanded in vitro. The human ADA (hADA) gene was introduced into these replicating ADA- T cells with the use of an amphotropic recombinant retrovirus carrying the hADA gene. Subsequently, infected T cells were selected on the basis of their ADA expression, by exposure to a combination of the toxic agent xylofuranosyl-adenine and the specific ADA inhibitor 2'-deoxycoformycin. CD4+ and CD8+ T cells could be infected and selected with equal efficiencies. The genetically modified T cells were shown to contain intact copies of the provirus and to express normal levels of hADA. As expected, uninfected T cells from the ADA-deficient patient displayed an increased sensitivity to 2'-deoxyadenosine. Following genetic modification, however, this sensitivity was restored to normal levels in both CD4+ and CD8+ T cells. The introduction of the hADA gene into the genome of the in vitro expanded T cells did not alter their phenotype, proliferative capacity or cytotoxic potential. These characteristics were identical to those of T cells derived from healthy individuals. These findings are of critical importance for the clinical application of hADA gene-transducted T cells.
authors.affiliation: Department of Immunology, Dr. Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: Braakman
authors.forename: V W
authors.initials: VW
authors.lastname: Van Beusechem
authors.forename: B A
authors.initials: BA
authors.lastname: Van Krimpen
authors.forename: A
authors.initials: A
authors.lastname: Fischer
authors.forename: R L
authors.initials: RL
authors.lastname: Bolhuis
authors.forename: D
authors.initials: D
authors.lastname: Valerio
mesh_terms: D000243:Adenosine Deaminase / Q000172:deficiency* / Q000235:genetics; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D002478:Cells, Cultured; D015316:Genetic Therapy; D006801:Humans; D007223:Infant; D010641:Phenotype; D013601:T-Lymphocytes / Q000201:enzymology*; D014161:Transduction, Genetic
chemical_list: D015704:CD4 Antigens; D016827:CD8 Antigens; D000243:Adenosine Deaminase