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.
| Original language | English |
|---|---|
| Pages (from-to) | 1096-1108 |
| Number of pages | 13 |
| Journal | Molecular Therapy |
| Volume | 10 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Dec 2004 |
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Keywords
- ADA-SCID
- Human stem/progenitor cells
- Immunodeficiency
- Lymphoid differentiation
ASJC Scopus subject areas
- Molecular Biology
Cite this
IL-3 or IL-7 increases ex vivo gene transfer efficiency in ADA-SCID BM CD34+ cells while maintaining in vivo lymphoid potential. / Ficara, Francesca; Superchi, Daniela B.; Hernández, Raisa Jofra; Mocchetti, Cristina; Carballido-Perrig, Nicole; Andolfi, Grazia; Deola, Sara; Colombo, Augusto; Bordignon, Claudio; Carballido, José M.; Roncarolo, Maria Grazia; Aiuti, Alessandro.
In: Molecular Therapy, Vol. 10, No. 6, 12.2004, p. 1096-1108.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - IL-3 or IL-7 increases ex vivo gene transfer efficiency in ADA-SCID BM CD34+ cells while maintaining in vivo lymphoid potential
AU - Ficara, Francesca
AU - Superchi, Daniela B.
AU - Hernández, Raisa Jofra
AU - Mocchetti, Cristina
AU - Carballido-Perrig, Nicole
AU - Andolfi, Grazia
AU - Deola, Sara
AU - Colombo, Augusto
AU - Bordignon, Claudio
AU - Carballido, José M.
AU - Roncarolo, Maria Grazia
AU - Aiuti, Alessandro
PY - 2004/12
Y1 - 2004/12
N2 - 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.
AB - 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.
KW - ADA-SCID
KW - Human stem/progenitor cells
KW - Immunodeficiency
KW - Lymphoid differentiation
UR - http://www.scopus.com/inward/record.url?scp=10344243971&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=10344243971&partnerID=8YFLogxK
U2 - 10.1016/j.ymthe.2004.08.014
DO - 10.1016/j.ymthe.2004.08.014
M3 - Article
C2 - 15564141
AN - SCOPUS:10344243971
VL - 10
SP - 1096
EP - 1108
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 6
ER -