Mobilized blood CD34+ cells transduced and selected with a clinically applicable protocol reconstitute lymphopoiesis in SCID-hu mice

Sara Deola, Samantha Scaramuzza, Roberto Sciarretta Birolo, Nicole Carballido-Perrig, Francesca Ficara, Cristina Mocchetti, Jonathan Dando, José M. Carballido, Claudio Bordignon, Maria Grazia Roncarolo, Marco Bregni, Alessandro Aiuti

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We developed a clinically applicable gene transfer procedure into mobilized peripheral blood (MPB) CD34+ hematopoietic progenitor cells, based on single viral exposure and selection of engineered cells. CD34+ cells were transduced with a retroviral vector carrying the truncated form of the nerve growth factor receptor (ΔNGFR) marker gene, and immunoselected for ΔNGFR expression. Optimal time and procedure for viral exposure, length of culture, and transgene expression of MPB CD34 + cells were determined using in vitro assays. The multipotent capacity of MPB CD34+-transduced cells was demonstrated in the SCID-hu bone/liver/thymus mouse model. Transduced ΔNGFR+ cells retained 50% of long-term culture-colony forming cells (LTC-CFC) compared to unmanipulated CD34+ cells. In SCID-hu mice, 52% of CD45+ cells, 27% of CD34+ cells, 49% of B cells, and more than 50% of T cells were derived from transplanted CD34+/ΔNGFR+ cells. Furthermore, transplantation of purified transduced cells greatly reduced the competition with untransduced progenitors occurring in unselected grafts. These data demonstrate that MPB CD34+ cells, transduced with a single viral exposure and selected by transgene expression, retain multilineage reconstitution capacity and remarkable transgene expression.

Original languageEnglish
Pages (from-to)305-311
Number of pages7
JournalHuman Gene Therapy
Volume15
Issue number3
DOIs
Publication statusPublished - 1 Mar 2004
Externally publishedYes

    Fingerprint

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Deola, S., Scaramuzza, S., Birolo, R. S., Carballido-Perrig, N., Ficara, F., Mocchetti, C., Dando, J., Carballido, J. M., Bordignon, C., Roncarolo, M. G., Bregni, M., & Aiuti, A. (2004). Mobilized blood CD34+ cells transduced and selected with a clinically applicable protocol reconstitute lymphopoiesis in SCID-hu mice. Human Gene Therapy, 15(3), 305-311. https://doi.org/10.1089/104303404322886156