title: Murine hematopoietic reconstitution after tagging and selection of retrovirally transduced bone marrow cells.
abstract: A major problem facing the effective treatment of patients with cancer is how to get the specific antitumor agent into every tumor cell. In this report we describe the use of a strategy that, by using retroviral vectors encoding a truncated human CD5 cDNA, allows the selection of only the infected cells, and we show the ability to obtain, before bone marrow transplantation, a population of 5-fluouracil-treated murine bone marrow cells that are 100% marked. This marked population of bone marrow cells is able to reconstitute the hematopoietic system in lethally irradiated mice, indicating that the surface marker lacks deleterious effects on the functionality of bone marrow cells. No gross abnormalities in hematopoiesis were detected in mice repopulated with CD5-expressing cells. Nevertheless, a significant proportion of the hematopoietic cells no longer expresses the surface marker CD5 in the 9-month-old recipient mice. This transcriptional inactivity of the proviral long terminal repeat (LTR) was accompanied by de novo methylation of the proviral sequences. Our results show that the use of the CD5 as a retrovirally encoded marker enables the rapid, efficient, and nontoxic selection in vitro of infected primary cells, which can entirely reconstitute the hematopoietic system in mice. These results should now greatly enhance the power of studies aimed at addressing questions such as generation of cancer-negative hematopoiesis.
authors.affiliation: Departamento de Proliferación y Diferenciación Celular, Instituto de Microbiología Bioquímica, Salamanca, Spain.
authors.forename: B
authors.initials: B
authors.lastname: García-Hernández
authors.forename: A
authors.initials: A
authors.lastname: Castellanos
authors.forename: A
authors.initials: A
authors.lastname: López
authors.forename: A
authors.initials: A
authors.lastname: Orfao
authors.forename: I
authors.initials: I
authors.lastname: Sánchez-García
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D018956:CD5 Antigens / Q000235:genetics; D002460:Cell Line; D002469:Cell Separation; D019175:DNA Methylation; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors; D006410:Hematopoiesis; D051379:Mice; D008810:Mice, Inbred C57BL; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D018956:CD5 Antigens