title: IL-3 or IL-7 increases ex vivo gene transfer efficiency in ADA-SCID BM CD34+ cells while maintaining in vivo lymphoid potential.
abstract: To improve maintenance and gene transfer of human lymphoid progenitors for clinical use in gene therapy of adenosine deaminase (ADA)-deficient SCID we investigated several gene transfer protocols using various stem cell-enriched sources. The lymphoid differentiation potential was measured by an in vitro clonal assay for B/NK cells and in the in vivo SCID-hu mouse model. Ex vivo culture with the cytokines TPO, FLT3-ligand, and SCF (T/F/S) plus IL-3 or IL-7 substantially increased the yield of transduced bone marrow (BM) CD34(+) cells purified from ADA-SCID patients or healthy donors, compared to T/F/S alone. Moreover, the use of IL-3 or IL-7 significantly improved the maintenance of in vitro B cell progenitors from ADA-SCID BM cells and allowed the efficient transduction of B and NK cell progenitors. Under these optimized conditions transduced CD34(+) cells were efficiently engrafted into SCID-hu mice and gave rise to B and T cell progeny, demonstrating the maintenance of in vivo lymphoid reconstitution capacity. The protocol based on the T/F/S + IL-3 combination was included in a gene therapy clinical trial for ADA-SCID, resulting in long-term engraftment of stem/progenitor cells. Remarkably, gene-corrected BM CD34(+) cells obtained from one patient 4 and 11 months after gene therapy were capable of repopulating the lymphoid compartment of SCID-hu hosts.
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy, 20132 Milan, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Ficara
authors.forename: Daniela B
authors.initials: DB
authors.lastname: Superchi
authors.forename: Raisa Jofra
authors.initials: RJ
authors.lastname: Hernández
authors.forename: Cristina
authors.initials: C
authors.lastname: Mocchetti
authors.forename: Nicole
authors.initials: N
authors.lastname: Carballido-Perrig
authors.forename: Grazia
authors.initials: G
authors.lastname: Andolfi
authors.forename: Sara
authors.initials: S
authors.lastname: Deola
authors.forename: Augusto
authors.initials: A
authors.lastname: Colombo
authors.forename: Claudio
authors.initials: C
authors.lastname: Bordignon
authors.forename: José M
authors.initials: JM
authors.lastname: Carballido
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Roncarolo
authors.forename: Alessandro
authors.initials: A
authors.lastname: Aiuti
mesh_terms: D000243:Adenosine Deaminase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D005312:Fetal Blood / Q000187:drug effects / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology*; D015851:Interleukin-7 / Q000494:pharmacology*; D008214:Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D016511:Severe Combined Immunodeficiency / Q000235:genetics / Q000378:metabolism / Q000473:pathology* / Q000628:therapy; D033581:Stem Cell Transplantation; D014161:Transduction, Genetic
chemical_list: D018952:Antigens, CD34; D007377:Interleukin-3; D015851:Interleukin-7; D000243:Adenosine Deaminase