Up-regulation of amphotrophic retroviral receptor expression in human peripheral blood CD34+ cells.
Retroviral-mediated gene transfer into hematopoietic stem cells provides the only means of stable transduction of these cells and their progeny for use with a variety of potentially therapeutic genes. Expression of the Moloney amphotropic retroviral receptor-pit-2 or GLVR-2-is critical to the recognition and entry of Moloney leukemia virus-derived viruses into human target cells such as CD34+ hematopoietic cells. GLVR-2 functions as a sodium-dependent phosphate transporter as well as a receptor. We have previously shown that the expression of the murine homologue of the amphotropic receptor Ram 1, also a phosphate transporter, is developmentally regulated in murine hematopoietic fetal liver cells. We also demonstrated that culture of murine fetal liver cells in phosphate-free (PO(4)-free) medium increases levels of receptor mRNA and makes murine fetal liver cells susceptible to Moloney amphotropic viral gene transfer. We now examine the effect of culture conditions on the expression of GLVR-2 in human CD34+ cells. In this report, we demonstrate that there is a 2-3 fold increase in GLVR-2 mRNA levels in CD34+ cells after 3 days in culture with interleukin 3, interleukin 6, and stem-cell factor. In addition, the use of PO(4)-free medium increases expression of GLVR-2 an additional 2-fold in these cells during this time. These results indicate that GLVR-2 expression can be up-regulated on these cells, and may permit improved retroviral gene transfer efficiencies.
Kaubisch
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A
Department of Medicine, Columbia University, College of Physicians and Surgeons, New York, New York 10032, USA.
Ward
M
M
Schoetz
S
S
Hesdorffer
C
C
Bank
A
A
D000818:Animals; D018952:Antigens, CD34; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000821:virology*; D006801:Humans; D008099:Liver / Q000166:cytology; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics; D028061:Phosphate Transport Proteins; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D011991:Receptors, Virus / Q000235:genetics*; D050608:Sodium-Phosphate Cotransporter Proteins; D050611:Sodium-Phosphate Cotransporter Proteins, Type III; D027981:Symporters; D015854:Up-Regulation
D018952:Antigens, CD34; D028061:Phosphate Transport Proteins; D012333:RNA, Messenger; D011991:Receptors, Virus; C493565:Slc20a2 protein, mouse; D050608:Sodium-Phosphate Cotransporter Proteins; D050611:Sodium-Phosphate Cotransporter Proteins, Type III; D027981:Symporters