IL-3 or IL-7 increases ex vivo gene transfer efficiency in ADA-SCID BM CD34+ cells while maintaining in vivo lymphoid potential

Francesca Ficara, Daniela B. Superchi, Raisa Jofra Hernández, Cristina Mocchetti, Nicole Carballido-Perrig, Grazia Andolfi, Sara Deola, Augusto Colombo, Claudio Bordignon, José M. Carballido, Maria Grazia Roncarolo, Alessandro Aiuti

Research output: Contribution to journalArticle

16 Citations (Scopus)

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 languageEnglish
Pages (from-to)1096-1108
Number of pages13
JournalMolecular Therapy
Volume10
Issue number6
DOIs
Publication statusPublished - 1 Dec 2004
Externally publishedYes

    Fingerprint

Keywords

  • ADA-SCID
  • Human stem/progenitor cells
  • Immunodeficiency
  • Lymphoid differentiation

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

Cite this

Ficara, F., Superchi, D. B., Hernández, R. J., Mocchetti, C., Carballido-Perrig, N., Andolfi, G., Deola, S., Colombo, A., Bordignon, C., Carballido, J. M., Roncarolo, M. G., & Aiuti, A. (2004). IL-3 or IL-7 increases ex vivo gene transfer efficiency in ADA-SCID BM CD34+ cells while maintaining in vivo lymphoid potential. Molecular Therapy, 10(6), 1096-1108. https://doi.org/10.1016/j.ymthe.2004.08.014