Successful transduction of human multipotent, lymphoid (T, B, NK) and myeloid, and transplantable CD34+CD38low cord blood cells using a murine oncoretroviral vector.
Hematopoietic stem cells (HSC) are subject to great interest because of their medical importance and their biological properties. Therefore, the possibility of genetically modifying human HSC is a major concern in several inherited pathologies. In this study, we aimed to demonstrate that a murine oncoretroviral vector can transduce multipotential cord blood (CB) stem cells. Sorted CB CD34(+)CD38(low) cells were transduced with a Moloney-based MFG retroviral vector containing the coding sequence of the murine CD2 (mCD2). CD34(+)mCD2(+) cells were sorted by flow cytometry and cultured either in bulk or at one cell per well in culture conditions that allow differentiation along lymphoid (T, B, and NK) and myeloid (M) lineages. Phenotypic analysis of cells generated in culture showed that CD34(+)mCD2(+) cells could give rise to all lymphoid and myeloid progeny, indicating that the MFG/mCD2 vector had transduced progenitors of all tested lineages. Moreover, clonal cultures of 660 CD34(+)mCD2(+) cells showed that approximately 5% of these cells were able to generate both myeloid and lymphoid (B + NK) progenies; for 25% of them, this included the production of lymphoid T cells. We also demonstrate that transduced CD34(+)CD38(low) CB cells with lymphoid and myeloid potentials were capable of engraftment into the bone marrow (BM) of nonobese diabetic-severe combined immunodeficiency (NOD-SCID) mice during several months. These results show that MFG retroviral vectors can transduce multipotent (T, B, NK, M) human hematopoietic progenitors with in vivo repopulating activity.
Ravet
E
E
INSERM U474, Maternité Port Royal, 123 Bd de Port Royal, 75014 Paris, France.
Dubart-Kupperschmitt
A
A
Robin
C
C
Titeux
M
M
Coulombel
L
L
Pflumio
F
F
D000818:Animals; D018952:Antigens, CD34; D001854:Bone Marrow Cells; D018801:CD2 Antigens / Q000235:genetics; D002465:Cell Movement; D036101:Cord Blood Stem Cell Transplantation / Q000379:methods*; D017930:Genes, Reporter; D005822:Genetic Vectors; D006085:Graft Survival; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D039902:Multipotent Stem Cells / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods* / Q000592:standards
D018952:Antigens, CD34; D018801:CD2 Antigens