title: Human adenosine deaminase expression in mice.
abstract: A replication defective retroviral vector containing a human adenosine deaminase (hADA) cDNA was produced by GP + E-86 packaging cells at high titer. We report long-term expression of hADA in the hematopoietic tissues of mice transplanted with bone marrow cells infected by in vitro co-cultivation with vector producing cells. Western analysis using an hADA-specific antibody allowed detection of the protein in the peripheral blood of all 37 transplanted mice for at least 9 weeks. Sixty-eight percent of the animals continued to express hADA in one or more of their hematopoietic tissues for the experimental period, and hADA was found in both spleen colonies and tissues of secondary recipients. There was provirus integration and expression in myeloid, erythroid, and lymphoid cell lineages, indicating extensive repopulation by the progeny of infected stem cells. The vector did not contain a selectable marker, and the infected stem cells did not have a competitive in vivo advantage. Nevertheless, we observed consistent gene transfer into hematopoietic stem cells and long-term expression of a human gene product in their progeny.
authors.affiliation: Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
authors.forename: K A
authors.initials: KA
authors.lastname: Moore
authors.forename: F A
authors.initials: FA
authors.lastname: Fletcher
authors.forename: D K
authors.initials: DK
authors.lastname: Villalon
authors.forename: A E
authors.initials: AE
authors.lastname: Utter
authors.forename: J W
authors.initials: JW
authors.lastname: Belmont
mesh_terms: D000243:Adenosine Deaminase / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D001773:Blood Cells / Q000201:enzymology; D015139:Blotting, Southern; D001853:Bone Marrow / Q000201:enzymology; D002455:Cell Division; D002460:Cell Line; D004247:DNA / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D009700:Nucleoside Deaminases / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012190:Retroviridae; D013154:Spleen / Q000166:cytology
chemical_list: D000906:Antibodies; D004247:DNA; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase