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.

  • Original Article
  • Published:

Cell-specific and efficient expression in mouse and human B cells by a novel hybrid immunoglobulin promoter in a lentiviral vector

Gene Therapy volume 14, pages 1623–1631 (2007)Cite this article

Abstract

The expression of genes specifically in B cells is of great interest in both experimental immunology as well as in future clinical gene therapy. We have constructed a novel enhanced B cell-specific promoter (Igk-E) consisting of an immunoglobulin kappa (Igk) minimal promoter combined with an intronic enhancer sequence and a 3′ enhancer sequence from Ig genes. The Igk-E promoter was cloned into a lentiviral vector and used to control expression of enhanced green fluorescent protein (eGFP). Transduction of murine B-cell lymphoma cell lines and activated primary splenic B cells, with IgK-E-eGFP lentivirus, resulted in expression of eGFP, as analysed by flow cytometry, whereas expression in non-B cells was absent. The specificity of the promoter was further examined by transducing Lin− bone marrow with Igk-E-eGFP lentivirus and reconstituting lethally irradiated mice. After 16 weeks flow cytometry of lymphoid tissues revealed eGFP expression by CD19+ cells, but not by CD3+, CD11b+, CD11c+ or Gr-1+ cells. CD19+ cells were comprised of both marginal zone B cells and recirculating follicular B cells. Activated human peripheral mononuclear cells were also transduced with Igk-E-eGFP lentivirus under conditions of selective B-cell activation. The Igk-E promoter was able to drive expression of eGFP only in CD19+ cells, while eGFP was expressed by both spleen focus-forming virus and cytomegalovirus constitutive promoters in CD19+ and CD3+ lymphocytes. These data demonstrate that in these conditions the Igk-E promoter is cell specific and controls efficient expression of a reporter protein in mouse and human B cells in the context of a lentiviral vector.

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
Figure 4

Similar content being viewed by others

References

  1. Cunningham-Rundles C, Ponda PP . Molecular defects in T- and B-cell primary immunodeficiency diseases. Nat Rev Immunol 2005; 5: 880–892.

    Article  CAS  Google Scholar 

  2. Gaspar HB, Howe S, Thrasher AJ . Gene therapy progress and prospects: gene therapy for severe combined immunodeficiency. Gene Therapy 2003; 10: 1999–2004.

    Article  CAS  Google Scholar 

  3. Larsen SR, Rasko JE . Lymphoproliferative disorders: prospects for gene therapy. Pathology 2005; 37: 523–533.

    Article  CAS  Google Scholar 

  4. Melo ME, El Amine M, Tonnetti L, Fleischman L, Scott DW . Gene therapeutic approaches to induction and maintenance of tolerance. Int Rev Immunol 2001; 20: 627–645.

    Article  CAS  Google Scholar 

  5. Wong W, Wood KJ . Transplantation tolerance by donor MHC gene transfer. Curr Gene Ther 2004; 4: 329–336.

    Article  CAS  Google Scholar 

  6. Laddy DJ, Weiner DB . From plasmids to protection: a review of DNA vaccines against infectious diseases. Int Rev Immunol 2006; 25: 99–123.

    Article  CAS  Google Scholar 

  7. Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996; 272: 263–267.

    Article  CAS  Google Scholar 

  8. Humeau LM, Binder GK, Lu X, Slepushkin V, Merling R, Echeagaray P et al. Efficient lentiviral vector-mediated control of HIV-1 replication in CD4 lymphocytes from diverse HIV+ infected patients grouped according to CD4 count and viral load. Mol Ther 2004; 9: 902–913.

    Article  CAS  Google Scholar 

  9. Bank A, Dorazio R, Leboulch P . A phase I/II clinical trial of beta-globin gene therapy for beta-thalassemia. Ann NY Acad Sci 2005; 1054: 308–316.

    Article  CAS  Google Scholar 

  10. Pawliuk R, Westerman KA, Fabry ME, Payen E, Tighe R, Bouhassira EE et al. Correction of sickle cell disease in transgenic mouse models by gene therapy. Science 2001; 294: 2368–2371.

    Article  CAS  Google Scholar 

  11. Indraccolo S, Minuzzo S, Roccaforte F, Zamarchi R, Habeler W, Stievano L et al. Effects of CD2 locus control region sequences on gene expression by retroviral and lentiviral vectors. Blood 2001; 98: 3607–3617.

    Article  CAS  Google Scholar 

  12. Marodon G, Mouly E, Blair EJ, Frisen C, Lemoine FM, Klatzmann D . Specific transgene expression in human and mouse CD4+ cells using lentiviral vectors with regulatory sequences from the CD4 gene. Blood 2003; 101: 3416–3423.

    Article  CAS  Google Scholar 

  13. Cui Y, Golob J, Kelleher E, Ye Z, Pardoll D, Cheng L . Targeting transgene expression to antigen-presenting cells derived from lentivirus-transduced engrafting human hematopoietic stem/progenitor cells. Blood 2002; 99: 399–408.

    Article  CAS  Google Scholar 

  14. Lutzko C, Senadheera D, Skelton D, Petersen D, Kohn DB . Lentivirus vectors incorporating the immunoglobulin heavy chain enhancer and matrix attachment regions provide position-independent expression in B lymphocytes. J Virol 2003; 77: 7341–7351.

    Article  CAS  Google Scholar 

  15. Werner M, Kraunus J, Baum C, Brocker T . B-cell-specific transgene expression using a self-inactivating retroviral vector with human CD19 promoter and viral post-transcriptional regulatory element. Gene Therapy 2004; 11: 992–1000.

    Article  CAS  Google Scholar 

  16. Moreau T, Bardin F, Imbert J, Chabannon C, Tonnelle C . Restriction of transgene expression to the B-lymphoid progeny of human lentivirally transduced CD34+ cells. Mol Ther 2004; 10: 45–56.

    Article  CAS  Google Scholar 

  17. Kalberer CP, Reininger L, Melchers F, Rolink AG . Priming of helper T cell-dependent antibody responses by hemagglutinin- transgenic B cells. Eur J Immunol 1997; 27: 2400–2407.

    Article  CAS  Google Scholar 

  18. Blankenstein T, Winter E, Muller W . A retroviral expression vector containing murine immunoglobulin heavy chain promoter/enhancer. Nucleic Acids Res 1988; 16: 10939.

    Article  CAS  Google Scholar 

  19. Bergman Y, Rice D, Grosschedl R, Baltimore D . Two regulatory elements for immunoglobulin kappa light chain gene expression. Proc Natl Acad Sci USA 1984; 81: 7041–7045.

    Article  CAS  Google Scholar 

  20. Pettersson S, Cook GP, Bruggemann M, Williams GT, Neuberger MS . A second B cell-specific enhancer 3′ of the immunoglobulin heavy-chain locus. Nature 1990; 344: 165–168.

    Article  CAS  Google Scholar 

  21. Cook GP, Meyer KB, Neuberger MS, Pettersson S . Regulated activity of the IgH intron enhancer (E mu) in the T lymphocyte lineage. Int Immunol 1995; 7: 89–95.

    Article  CAS  Google Scholar 

  22. Demaison C, Parsley K, Brouns G, Scherr M, Battmer K, Kinnon C et al. High-level transduction and gene expression in hematopoietic repopulating cells using a human imunodeficiency virus type 1-based lentiviral vector containing an internal spleen focus forming virus promoter. Hum Gene Ther 2002; 13: 803–813.

    Article  CAS  Google Scholar 

  23. Janssens W, Chuah MK, Naldini L, Follenzi A, Collen D, Saint-Remy JM et al. Efficiency of onco-retroviral and lentiviral gene transfer into primary mouse and human B-lymphocytes is pseudotype dependent. Hum Gene Ther 2003; 14: 263–276.

    Article  CAS  Google Scholar 

  24. Rossi GR, Mautino MR, Morgan RA . High-efficiency lentiviral vector-mediated gene transfer into murine macrophages and activated splenic B lymphocytes. Hum Gene Ther 2003; 14: 385–391.

    Article  CAS  Google Scholar 

  25. Warncke M, Vogt B, Ulrich J, von Laer MD, Beyer W, Klump H et al. Efficient in vitro transduction of naive murine B cells with lentiviral vectors. Biochem Biophys Res Commun 2004; 318: 673–679.

    Article  CAS  Google Scholar 

  26. Cooper MD . Pre-B cells; normal and abnormal development. J Clin Immunol 1981; 1: 81–89.

    Article  CAS  Google Scholar 

  27. Chinnasamy D, Chinnasamy N, Enriquez MJ, Otsu M, Morgan RA, Candotti F . Lentiviral-mediated gene transfer into human lymphocytes: role of HIV-1 accessory proteins. Blood 2000; 96: 1309–1316.

    CAS  PubMed  Google Scholar 

  28. Cavazzana-Calvo M, Hacein-Bey S, de Saint BG, Gross F, Yvon E, Nusbaum P et al. Gene therapy of human severe combined immunodeficiency (SCID)-X1 disease. Science 2000; 288: 669–672.

    Article  CAS  Google Scholar 

  29. Gaspar HB, Parsley KL, Howe S, King D, Gilmour KC, Sinclair J et al. Gene therapy of X-linked severe combined immunodeficiency by use of a pseudotyped gammaretroviral vector. Lancet 2004; 364: 2181–2187.

    Article  CAS  Google Scholar 

  30. Kim KJ, Kanellopoulos-Langevin C, Merwin RM, Sachs DH, Asofsky R . Establishment and characterization of BALB/c lymphoma lines with B cell properties. J Immunol 1979; 122: 549–554.

    CAS  PubMed  Google Scholar 

  31. Aaronson SA, Todaro GJ . Development of 3T3-like lines from Balb-c mouse embryo cultures: transformation susceptibility to SV40. J Cell Physiol 1968; 72: 141–148.

    Article  CAS  Google Scholar 

  32. Corthay A, Backlund J, Broddefalk J, Michaelsson E, Goldschmidt TJ, Kihlberg J et al. Epitope glycosylation plays a critical role for T cell recognition of type II collagen in collagen-induced arthritis. Eur J Immunol 1998; 28: 2580–2590.

    Article  CAS  Google Scholar 

  33. Williams RL, Courtneidge SA, Wagner EF . Embryonic lethalities and endothelial tumors in chimeric mice expressing polyoma virus middle T oncogene. Cell 1988; 52: 121–131.

    Article  CAS  Google Scholar 

  34. Zufferey R, Nagy D, Mandel RJ, Naldini L, Trono D . Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat Biotechnol 1997; 15: 871–875.

    Article  CAS  Google Scholar 

  35. Neil S, Martin F, Ikeda Y, Collins M . Postentry restriction to human immunodeficiency virus-based vector transduction in human monocytes. J Virol 2001; 75: 5448–5456.

    Article  CAS  Google Scholar 

  36. Laurie KL, van Driel IR, Zwar TD, Barrett SP, Gleeson PA . Endogenous H/K ATPase beta-subunit promotes T cell tolerance to the immunodominant gastritogenic determinant. J Immunol 2002; 169: 2361–2367.

    Article  CAS  Google Scholar 

  37. Charrier S, Stockholm D, Seye K, Opolon P, Taveau M, Gross DA et al. A lentiviral vector encoding the human Wiskott–Aldrich syndrome protein corrects immune and cytoskeletal defects in WASP knockout mice. Gene Therapy 2005; 12: 597–606.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the Plasmid Factory (Bielefeld, Germany) for production of plasmids pCMVΔR8.91 and pMD.G2. These studies were supported by the Arthritis Research Campaign (KG), Wellcome Trust (AJT), Leukemia Research Fund LRF (WQ) and EU grant 6th Framework (CONSERT) (SH).

Author information

Authors and Affiliations

  1. Molecular Immunology Unit, Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, London, UK

    K L Laurie, M P Blundell, S J Howe, J Sinclair, W Qasim, A J Thrasher & K Gustafsson

  2. Infectious Diseases and Microbiology Unit, UCL Institute of Child Health, London, UK

    H E Baxendale

  3. Department of Experimental Medical Science, Section for Medical Inflammation Research, Lund University, Lund, Sweden

    U Brunsberg & R Holmdahl

  4. Great Ormond Street Hospital for Children NHS Trust, London, UK

    A J Thrasher

Authors
  1. K L Laurie
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M P Blundell
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H E Baxendale
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S J Howe
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J Sinclair
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. W Qasim
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. U Brunsberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. A J Thrasher
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. R Holmdahl
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. K Gustafsson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K Gustafsson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Laurie, K., Blundell, M., Baxendale, H. et al. Cell-specific and efficient expression in mouse and human B cells by a novel hybrid immunoglobulin promoter in a lentiviral vector. Gene Ther 14, 1623–1631 (2007). https://doi.org/10.1038/sj.gt.3303021

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3303021

Keywords

This article is cited by

Search

Advanced search

Quick links