I'll do the CEGE post while I'm here...
Here is the press release...which is a
pretty good summary already...
Cell Genesys Reports Efficient Gene Transfer to
Bone Marrow Stem Cells With Lentiviral Gene Therapy Technology
biz.yahoo.com
...and here is the abstract paste from PNAS:
Vol. 96, Issue 6, 2988-2993, March 16, 1999
Immunology
Stable transduction of quiescent CD34+CD38 human hematopoietic cells by HIV-1-based lentiviral vectors
Scott S. Case*, Mary A. Price*, Craig T. Jordan, Xiao Jin Yu*, Lijun Wang*, Gerhard Bauer*, Dennis L. Haas*, Dakun Xu*, Renata Stripecke*, Luigi Naldini, Donald B. Kohn*, and Gay M. Crooks*,§
* Division of Research Immunology/Bone Marrow Transplantation, Childrens Hospital Los Angeles, Los Angeles, CA 90027; Blood and Marrow Transplant Program, Markey Cancer Center, University of Kentucky, Lexington, KY 40536-0093; and Cell Genesys, Foster City, CA 94404
Edited by Inder M. Verma, The Salk Institute for Biological Studies, San Diego, CA, and approved January 18, 1999 (received for review November 9, 1998)
We compared the efficiency of transduction by an HIV-1-based lentiviral vector to that by a Moloney murine leukemia virus (MLV) retroviral vector, using stringent in vitro assays of primitive, quiescent human hematopoietic progenitor cells. Each construct contained the enhanced green fluorescent protein (GFP) as a reporter gene. The lentiviral vector, but not the MLV vector, expressed GFP in nondivided CD34+ cells (45.5% GFP+) and in CD34+CD38 cells in G0 (12.4% GFP+), 48 hr after transduction. However, GFP could also be detected short-term in CD34+ cells transduced with a lentiviral vector that contained a mutated integrase gene. The level of stable transduction from integrated vector was determined after extended long-term bone marrow culture. Both MLV vectors and lentiviral vectors efficiently transduced cytokine-stimulated CD34+ cells. The MLV vector did not transduce more primitive, quiescent CD34+CD38 cells (n = 8). In contrast, stable transduction of CD34+CD38 cells by the lentiviral vector was seen for over 15 weeks of extended long-term culture (9.2 ± 5.2%, n = 7). GFP expression in clones from single CD34+CD38 cells confirmed efficient, stable lentiviral transduction in 29% of early and late-proliferating cells. In the absence of growth factors during transduction, only the lentiviral vector was able to transduce CD34+ and CD34+CD38 cells (13.5 ± 2.5%, n = 11 and 12.2 ± 9.7%, n = 4, respectively). The lentiviral vector is clearly superior to the MLV vector for transduction of quiescent, primitive human hematopoietic progenitor cells and may provide therapeutically useful levels of gene transfer into human hematopoietic stem cells.
--------------------------------------------------------------------------------
§ To whom reprint requests should be addressed at: Childrens Hospital Los Angeles, 4650 Sunset Blvd., MS #62, Los Angeles, CA 90027. e-mail: gcrooks@chla.usc.edu.
Copyright © 1999 by The National Academy of Sciences
|
