Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Retrovirus-mediated WASP gene transfer corrects defective actin polymerization in B cell lines from Wiskott–Aldrich syndrome patients carrying ‘null’ mutations

Gene Therapy volume 6, pages 1170–1174 (1999)Cite this article

Abstract

Boys affected with Wiskott–Aldrich syndrome (WAS) present with variable association of thrombocytopenia, eczema and immune deficiency. If untreated, WAS patients may succumb to intracerebral hemorrhages, severe infections or malignancies. Allogeneic bone marrow transplantation (BMT) can cure all aspects of the disease, but HLA-identical donors are not available to all patients and mismatched BMTs are unfortunately associated with high mortality and morbidity. The good success of HLA-matched BMT, however, makes WAS a potential candidate for hematopoietic stem cell gene therapy. WAS patients carry mutations of the Wiskott–Aldrich syndrome protein gene encoding WASP, a 502-amino acid proline-rich protein with demonstrated involvement in the organization of the actin cytoskeleton. To verify the feasibility of genetic correction for this disease, the WASP cDNA was expressed in EBV-immortalized B cell lines obtained from WAS patients using a retroviral vector. Transduced WAS cells showed levels of WASP expression similar to those found in cells from normal donors, without detectable effects on viability or growth characteristics. In addition, retrovirus-mediated expression of WASP led to improvement of cytoplasmic F-actin expression and formation of F-actin-positive microvilli, a process shown to be defective in untransduced WAS cell lines. These preliminary results indicate a potential use for retrovirus-mediated gene transfer as therapy for WAS.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Sullivan KE, Mullen CA, Blaese RM, Winkelstein JA . A multiinstitutional survey of the Wiskott–Aldrich syndrome J Pediatr 1994 125: 876–885

    Article  CAS  PubMed  Google Scholar 

  2. Ochs HD . The Wiskott–Aldrich syndrome Springer Semin Immunopathol 1998 19: 435–458

    Article  CAS  PubMed  Google Scholar 

  3. Derry JM, Ochs HD, Francke U . Isolation of a novel gene mutated in Wiskott–Aldrich syndrome Cell 1994 78: 635–644

    Article  CAS  PubMed  Google Scholar 

  4. Villa A et al. X-linked thrombocytopenia and Wiskott–Aldrich syndrome are allelic diseases with mutations in the WASP gene Nat Genet 1995 9: 414–417

    Article  CAS  PubMed  Google Scholar 

  5. Stewart DM et al. Studies of the expression of the Wiskott–Aldrich syndrome protein J Clin Invest 1996 97: 2627–2634

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Parolini O et al. Expression of Wiskott–Aldrich syndrome protein (WASP) gene during hematopoietic differentiation Blood 1997 90: 70–75

    CAS  PubMed  Google Scholar 

  7. Aspenstrom P, Lindberg U, Hall A . Two GTPases, Cdc42 and Rac, bind directly to a protein implicated in the immunodeficiency disorder Wiskott–Aldrich syndrome Curr Biol 1996 6: 70–75

    Article  CAS  PubMed  Google Scholar 

  8. Symons M et al. Wiskott–Aldrich syndrome protein, a novel effector for the GTPase CDC42Hs, is implicated in actin polymerization Cell 1996 84: 723–734

    Article  CAS  PubMed  Google Scholar 

  9. Kolluri R et al. Direct interaction of the Wiskott Aldrich syndrome protein with the GTPase Cdc42 Proc Natl Acad Sci USA 1996 93: 5616–5618

    Article  Google Scholar 

  10. Ramesh N, Anton IM, Hartwig JH, Geha RS . WIP, a protein associated with Wiskott–Aldrich syndrome protein, induces actin polymerization and redistribution in lymphoid cells Proc Natl Acad Sci USA 1997 94: 14671–14676

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Brochstein JA et al. Marrow transplantation from human leukocyte antigen-identical or haploidentical donors for correction of Wiskott–Aldrich syndrome J Pediatr 1991 119: 907–912

    Article  CAS  PubMed  Google Scholar 

  12. Filipovich A . Stem cell transplantation from unrelated donors for correction of primary immunodeficiencies Immunol All Clin N Am 1996 16: 377–392

    Article  Google Scholar 

  13. Miller AD et al. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus J Virol 1991 65: 2220–2224

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Treisman J et al. Interleukin-2-transduced lymphocytes grow in an autocrine fashion and remain responsive to antigen Blood 1995 85: 139–145

    CAS  PubMed  Google Scholar 

  15. Bunnell BA et al. High-efficiency retrovirus-mediated gene transfer into human and nonhuman primate peripheral blood lymphocytes Proc Natl Acad Sci USA 1995 92: 7739–7743

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Kenney D et al. Morphological abnormalities in the lymphocytes of patients with the Wiskott–Aldrich syndrome Blood 1986 68: 1329–1332

    CAS  PubMed  Google Scholar 

  17. Gallego MD, Santamaria M, Pena J, Molina IJ . Defective actin reorganization and polymerization of Wiskott–Aldrich T cells in response to CD3-mediated stimulation Blood 1997 90: 3089–3097

    CAS  PubMed  Google Scholar 

  18. Facchetti F et al. Defective actin polymerization in EBV-transformed B-cell lines from patients with the Wiskott–Aldrich syndrome J Pathol 1998 185: 99–107

    Article  CAS  PubMed  Google Scholar 

  19. Snapper SB et al. Wiskott–Aldrich syndrome protein-deficient mice reveal a role for WASP in T but not B cell activation Immunity 1998 9: 81–91

    Article  CAS  PubMed  Google Scholar 

  20. Huang MM et al. Expression of human Wiskott–Aldrich-syndrome protein in patients primary hematopoietic-cells and Ebv-transduced cell-lines after retroviral vector-mediated gene transduction Blood 1997 90: 1154

    Google Scholar 

  21. Fischer A et al. Bone marrow transplantation (BMT) in Europe for primary immunodeficiencies other than severe combined immunodeficiency: a report from the European Group for BMT and the European Group for Immunodeficiency Blood 1994 83: 1149–1154

    CAS  PubMed  Google Scholar 

  22. Molina IJ et al. T cells of patients with the Wiskott–Aldrich syndrome have a restricted defect in proliferative responses J Immunol 1993 151: 4383–4390

    CAS  PubMed  Google Scholar 

  23. Simon HU, Mills GB, Hashimoto S, Siminovitch KA . Evidence for defective transmembrane signaling in B cells from patients with Wiskott–Aldrich syndrome J Clin Invest 1992 90: 1396–1405

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Henriquez NV, Rijkers GT, Zegers BJ . Antigen receptor-mediated transmembrane signaling in Wiskott–Aldrich syndrome J Immunol 1994 153: 395–399

    CAS  PubMed  Google Scholar 

  25. Tiberghien P et al. Ganciclovir treatment of herpes simplex thymidine kinase-transduced primary T lymphocytes: an approach for specific in vivo donor T-cell depletion after bone marrow transplantation? Blood 1994 84: 1333–1341

    CAS  PubMed  Google Scholar 

  26. Candotti F et al. Retroviral-mediated gene correction for X-linked severe combined immunodeficiency Blood 1996 87: 3097–3102

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr S Weissman for donating the WASP cDNA and to Prof LD Notarangelo for insightful discussion and continuous support.

Author information

Authors and Affiliations

  1. Clinical Gene Therapy Branch, National Human Genome Research Institute, NIH, Bethesda, MD, USA

    F Candotti & R M Blaese

  2. Department of Pathology, University of Brescia, Spedali Civili, Brescia, Italy

    F Facchetti & L Blanzuoli

  3. Immunophysiology Section, Metabolism Branch, National Cancer Institute, NIH, Bethesda, MD, USA

    D M Stewart & D L Nelson

Authors
  1. F Candotti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F Facchetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L Blanzuoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D M Stewart
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D L Nelson
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R M Blaese
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Candotti, F., Facchetti, F., Blanzuoli, L. et al. Retrovirus-mediated WASP gene transfer corrects defective actin polymerization in B cell lines from Wiskott–Aldrich syndrome patients carrying ‘null’ mutations. Gene Ther 6, 1170–1174 (1999). https://doi.org/10.1038/sj.gt.3300926

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3300926

Keywords

This article is cited by

Search

Advanced search

Quick links