title: Transduction efficiency of cell lines and hematopoietic stem cells correlates with retrovirus receptor mRNA levels.
abstract: The low transduction efficiency of hematopoietic stem cells (HSC) using amphotropic retroviruses continues to plague gene therapy protocols. This low transduction efficiency may be related to a low level of amphotropic retrovirus binding to target cell receptors. We have assayed murine and human cell lines as well as primary bone marrow HSC populations for mRNA encoding retrovirus receptors. Total cellular RNA was amplified by reverse transcriptase-polymerase chain reaction and the level of ecotropic and amphotropic receptor mRNA was compared to the level of beta 2-microglobulin mRNA in the same cell populations. Cell lines that are easily transduced by ecotropic and amphotropic retroviruses have high levels of receptor mRNA. In studies using murine HSC-enriched populations obtained from bone marrow, we observed a high correlation between transduction efficiency and the level of ecotropic and amphotropic receptor mRNA. We predict from these findings that purification of monkey and human HSC populations with high levels of amphotropic receptor mRNA will enable us to obtain improved efficiency of gene transfer.
authors.affiliation: Laboratory of Gene Transfer, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: D
authors.initials: D
authors.lastname: Orlic
authors.forename: L J
authors.initials: LJ
authors.lastname: Girard
authors.forename: S M
authors.initials: SM
authors.lastname: Anderson
authors.forename: B K
authors.initials: BK
authors.lastname: Do
authors.forename: N E
authors.initials: NE
authors.lastname: Seidel
authors.forename: C T
authors.initials: CT
authors.lastname: Jordan
authors.forename: D M
authors.initials: DM
authors.lastname: Bodine
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006412:Hematopoietic Stem Cells / Q000737:chemistry* / Q000821:virology; D006801:Humans; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis*; D011991:Receptors, Virus / Q000235:genetics*; D012190:Retroviridae; D014170:Transformation, Genetic
chemical_list: D012333:RNA, Messenger; D011991:Receptors, Virus