Mobilization and transduction of CD34(+) peripheral blood stem cells in patients with X-linked chronic granulomatous disease.

As a single-gene defect in phagocytes, the X-linked form of chronic granulomatous disease (X-CGD) is a disorder potentially amenable to gene therapy by transfer of a functional copy of the gp91(phox) gene into hematopoietic stem cells (HSC). Although antimicrobial agents and interferon-gamma (IFN-gamma) have significantly improved its prognosis, CGD is still associated with high morbidity and mortality. The disease can be cured by bone marrow transplantation (BMT); however, BMT in CGD has been associated with unacceptably high rates of morbidity, mortality, and graft failure, except in very selected cases in which an HLA-identical donor is available. Prerequisites for a clinical gene therapy of CGD are an efficient mobilization of peripheral blood stem cells (PBSC) as well as the preservation of their viability and hematopoietic potential following transduction and ex vivo culture. We show that (i) mobilization and collection of CD34(+) cells after a 4-week IFN-gamma-free period by G-CSF results in sufficient numbers of cells for transplantation; (ii) the quality of collected stem cells is not altered in comparison to cells obtained from healthy volunteers as assessed by long-term culture initiating cells (LTC-IC) and progenitor cell expansion; (iii) retroviral transfer of the gp91(phox) gene under defined, serum-free conditions leads to high and stable reconstitution of the respiratory burst activity in X-CGD neutrophils derived from transduced CD34(+) progenitor and LTC-IC. Withdrawal of IFN-gamma in CGD patients may improve mobilization of CD34(+) stem cells by G-CSF. The gene transfer conditions established here are applicable to a clinical approach for gene therapy of X-CGD.

[1]  M. Dinauer,et al.  Variable correction of host defense following gene transfer and bone marrow transplantation in murine X-linked chronic granulomatous disease. , 2001, Blood.

[2]  H. Malech Use of Serum‐Free Medium with Fibronectin Fragment Enhanced Transduction in a System of Gas Permeable Plastic Containers to Achieve High Levels of Retrovirus Transduction at Clinical Scale , 2000, Stem cells.

[3]  R. Storb,et al.  The use of granulocyte colony-stimulating factor during retroviral transduction on fibronectin fragment CH-296 enhances gene transfer into hematopoietic repopulating cells in dogs. , 1999, Blood.

[4]  N. Young,et al.  Expression of interferon-gamma by stromal cells inhibits murine long-term repopulating hematopoietic stem cell activity. , 1999, Experimental hematology.

[5]  C. Eaves,et al.  Optimization of retroviral-mediated gene transfer to human NOD/SCID mouse repopulating cord blood cells through a systematic analysis of protocol variables. , 1999, Experimental hematology.

[6]  A. Ganser,et al.  Correction of respiratory burst activity in X-linked chronic granulomatous cells to therapeutically relevant levels after gene transfer into bone marrow CD34+ cells. , 1998, Human gene therapy.

[7]  W. Piacibello,et al.  Differential growth factor requirement of primitive cord blood hematopoietic stem cell for self-renewal and amplification vs proliferation and differentiation , 1998, Leukemia.

[8]  S. Holland,et al.  Prolonged production of NADPH oxidase-corrected granulocytes after gene therapy of chronic granulomatous disease. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[9]  H. Malech,et al.  Genetic correction of p67phox deficient chronic granulomatous disease using peripheral blood progenitor cells as a target for retrovirus mediated gene transfer. , 1997, Blood.

[10]  C. Hannum,et al.  FLT3 ligand preserves the ability of human CD34+ progenitors to sustain long-term hematopoiesis in immune-deficient mice after ex vivo retroviral-mediated transduction. , 1997, Blood.

[11]  M. Dinauer,et al.  Retroviral-mediated gene transfer of gp91phox into bone marrow cells rescues defect in host defense against Aspergillus fumigatus in murine X-linked chronic granulomatous disease. , 1997, Blood.

[12]  C. Eaves,et al.  Enhanced detection, maintenance, and differentiation of primitive human hematopoietic cells in cultures containing murine fibroblasts engineered to produce human steel factor, interleukin-3, and granulocyte colony-stimulating factor. , 1996, Blood.

[13]  S. Hegewisch-Becker,et al.  High-dose multidrug resistance in primary human hematopoietic progenitor cells transduced with optimized retroviral vectors. , 1996, Blood.

[14]  Jeffery L. Miller,et al.  Granulocyte colony-stimulating factor recruitment of CD34+ progenitors to peripheral blood : Impaired mobilization in chronic granulomatous disease and adenosine deaminase-deficient severe combined immunodeficiency disease patients , 1996 .

[15]  Dirk Roos,et al.  Mutations in the X-linked and autosomal recessive forms of chronic granulomatous disease , 1996 .

[16]  S. Hegewisch-Becker,et al.  Novel retroviral vectors for efficient expression of the multidrug resistance (mdr-1) gene in early hematopoietic cells , 1995, Journal of virology.

[17]  N. Young,et al.  Interferon-gamma and tumor necrosis factor-alpha suppress both early and late stages of hematopoiesis and induce programmed cell death. , 1995, Journal of cellular physiology.

[18]  R. Levinsky,et al.  Gene transfer to primary chronic granulomatous disease monocytes , 1995, The Lancet.

[19]  H. Snoeck,et al.  Interferon gamma selectively inhibits very primitive CD342+CD38- and not more mature CD34+CD38+ human hematopoietic progenitor cells , 1994, The Journal of experimental medicine.

[20]  E. Shpall,et al.  Transplantation of enriched CD34-positive autologous marrow into breast cancer patients following high-dose chemotherapy: influence of CD34-positive peripheral-blood progenitors and growth factors on engraftment. , 1994, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[21]  J. Garcia,et al.  Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus , 1991, Journal of virology.

[22]  W. Ostertag,et al.  Embryonic stem cell virus, a recombinant murine retrovirus with expression in embryonic stem cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[23]  A. Miller,et al.  Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection , 1990, Molecular and cellular biology.

[24]  C. Eaves,et al.  Functional characterization of individual human hematopoietic stem cells cultured at limiting dilution on supportive marrow stromal layers. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Curnutte,et al.  Kinetic microplate assay for superoxide production by neutrophils and other phagocytic cells. , 1990, Methods in enzymology.

[26]  P. Lansdorp,et al.  Characterization and partial purification of human marrow cells capable of initiating long-term hematopoiesis in vitro. , 1989, Blood.

[27]  G. Trinchieri,et al.  Effects of recombinant tumor necrosis factor, lymphotoxin, and immune interferon on proliferation and differentiation of enriched hematopoietic precursor cells. , 1988, Experimental hematology.