Long-term multilineage expression in peripheral blood from a Moloney murine leukemia virus vector after serial transplantation of transduced bone marrow cells.
Using a mouse bone marrow transplantation model, the authors evaluated a Moloney murine leukemia virus (MMLV)-based vector encoding 2 anti-human immunodeficiency virus genes for long-term expression in blood cells. The vector also encoded the human nerve growth factor receptor (NGFR) to serve as a cell-surface marker for in vivo tracking of transduced cells. NGFR(+) cells were detected in blood leukocytes of all mice (n=16; range 16%-45%) 4 to 5 weeks after transplantation and were repeatedly detected in blood erythrocytes, platelets, monocytes, granulocytes, T cells, and B cells of all mice for up to 8 months. Transgene expression in individual mice was not blocked in the various cell lineages of the peripheral blood and spleen, in several stages of T-cell maturation in the thymus, or in the Lin(-/lo)Sca-1(+) and c-kit(+)Sca-1(+) subsets of bone marrow cells highly enriched for long-term multilineage-reconstituting activity. Serial transplantation of purified NGFR(+)c-kit(+)Sca-1(+) bone marrow cells resulted in the reconstitution of multilineage hematopoiesis by donor type NGFR(+) cells in all engrafted mice. The authors concluded that MMLV-based vectors were capable of efficient and sustained transgene expression in multiple lineages of peripheral blood cells and hematopoietic organs and in hematopoietic stem cell (HSC) populations. Differentiation of engrafting HSC to peripheral blood cells is not necessarily associated with dramatic suppression of retroviral gene expression. In light of earlier studies showing that vector elements other than the long-terminal repeat enhancer, promoter, and primer binding site can have an impact on long-term transgene expression, these findings accentuate the importance of empirically testing retroviral vectors to determine lasting in vivo expression.
Austin
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SyStemix Inc, Palo Alto, CA 94304, USA. tim.austin@pharma.novartis.com
Salimi
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Veres
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Morel
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Ilves
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H
Scollay
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Plavec
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I
D000818:Animals; D001773:Blood Cells / Q000378:metabolism / Q000821:virology*; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation; D019070:Cell Lineage; D016373:DNA, Antisense / Q000235:genetics; D017930:Genes, Reporter; D015751:Genes, pol; D015757:Genes, rev; D005822:Genetic Vectors / Q000096:biosynthesis*; D006085:Graft Survival; D015497:HIV-1 / Q000235:genetics; D006410:Hematopoiesis; D051379:Mice; D008810:Mice, Inbred C57BL; D008979:Moloney murine leukemia virus / Q000235:genetics* / Q000502:physiology; D017475:Receptors, Nerve Growth Factor / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D013154:Spleen / Q000166:cytology / Q000821:virology; D013601:T-Lymphocytes / Q000378:metabolism / Q000821:virology; D013997:Time Factors; D014779:Virus Replication
D016373:DNA, Antisense; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins