Genetic correction of cultured T cells from an adenosine deaminase-deficient patient: characteristics of non-transduced and transduced T cells.
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.
Braakman
E
E
Department of Immunology, Dr. Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
Van Beusechem
V W
VW
Van Krimpen
B A
BA
Fischer
A
A
Bolhuis
R L
RL
Valerio
D
D
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
D015704:CD4 Antigens; D016827:CD8 Antigens; D000243:Adenosine Deaminase