Restriction of transgene expression to the B-lymphoid progeny of human lentivirally transduced CD34+ cells.
Development of gene transfer strategies will necessitate improved efficiency and control of transduction and transgene expression. We here provide evidence that targeting expression of the GFP reporter gene to the B-lymphoid progeny of genetically modified human hematopoietic progenitor cells can be achieved through the insertion of regulatory sequences from the human CD19 gene promoter into a lentiviral vector. Based on a bioinformatics approach, three human CD19-derived sequences were designed and inserted into a self-inactivated lentiviral vector backbone upstream of the GFP gene: S.CD19 (230 bp), M.CD19 (464 bp), and L.CD19 (1274 bp). These new lentiviral vectors efficiently transduced cord blood CD34(+) cells. The M.CD19 and especially L.CD19 sequences preferentially targeted GFP expression to in vitro and in vivo differentiated CD19(+) progeny; moreover, transgene expression was detected from the CD34(+) pro/pre-B cell to the mature peripheral IgM(+) B cell stage. In contrast, GFP expression was weak or absent in primary T-lymphoid and uncommitted progenitor cells or in erythroid, natural killer, or myeloid differentiated cells. Such B-lineage-specific lentiviral vectors may be useful for correcting inherited disorders that affect B-lymphoid cells or for deciphering the transcriptional program that controls B cell commitment and differentiation.
Moreau
Thomas
T
Centre de Thérapie Cellulaire et Génique, Institut Paoli-Calmettes, Centre Régional de Lutte contre le Cancer Provence-Alpes-Côte d'Azur, Marseille, France.
Bardin
Florence
F
Imbert
Jean
J
Chabannon
Christian
C
Tonnelle
Cécile
C
D000818:Animals; D018941:Antigens, CD19 / Q000235:genetics*; D018952:Antigens, CD34 / Q000032:analysis*; D001402:B-Lymphocytes / Q000378:metabolism* / Q000637:transplantation; D002460:Cell Line; D005312:Fetal Blood / Q000166:cytology; D015870:Gene Expression / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics; D014183:Transplantation, Heterologous
D018941:Antigens, CD19; D018952:Antigens, CD34; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins