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Genomics, Gene Therapy and Proteomics

Correction of B-cell development in Btk-deficient mice using lentiviral vectors with codon-optimized human BTK

Leukemia volume 24, pages 1617–1630 (2010)Cite this article

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Abstract

X-linked agammaglobulinemia (XLA) is the most common primary immunodeficiency (PID) in man and caused by mutations in the Bruton’s tyrosine kinase (BTK) gene. XLA is characterized by a B-cell differentiation arrest in bone marrow, absence of mature B cells and immunoglobulins (Igs), and recurrent bacterial infections. We used self-inactivating lentiviral vectors expressing codon-optimized human BTK under the control of three different ubiquitous or B cell-specific promoters. Btk−/− mice engrafted with transduced cells showed correction of both precursor B-cell and peripheral B-cell development. Lentiviral vectors containing the wildtype BTK sequence did not correct the phenotype. All treated mice with codon-optimized BTK exhibited the recovery of B1 cells in the peritoneal cavity, and of serum IgM and IgG3 levels. Calcium mobilization responses upon B-cell receptor stimulation as well as in vivo responses to T cell-independent antigens were restored. Viral promoters overexpressing BTK >100-fold above normal resulted in erythro-myeloid proliferations independent of insertional mutagenesis. However, transplantation into secondary Btk−/− recipients using cellular promoters resulted in functional restoration of peripheral B cells and IgM levels, without any adverse effects. In conclusion, transduction of human BTK corrects B-cell development and antigen-specific antibody responses in Btk−/− mice, thus indicating the feasibility of lentiviral gene therapy for XLA, provided that BTK expression does not vastly exceed normal levels.

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Acknowledgements

We would like to thank Tiago Luis, Brigitta Naber, Edwin de Hass and Machteld Tiemessen (Staal laboratory) for their technical assistance and Yvette Caljouw for her assistance at the EDC. This work was supported in part by a Grant from the Translational Gene Therapy Research Program of ZonMw—the Netherlands Organization for Health Research and Development (project no. 43100016).

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Authors and Affiliations

  1. Department of Immunology, Erasmus MC, Rotterdam, The Netherlands

    Y Y Ng, M R M Baert, K Pike-Overzet, M Rodijk, R W Hendriks, J J M van Dongen & F J T Staal

  2. Department of Immunohematology and Blood transfusion, Leiden University Medical Center, Leiden, The Netherlands

    Y Y Ng, M R M Baert, K Pike-Overzet, M Rodijk & F J T Staal

  3. Experimental Hematology, Hannover Medical School, Hannover, Germany

    M H Brugman, A Schambach & C Baum

  4. Department of Pulmonary Medicine, Erasmus MC, Rotterdam, The Netherlands

    R W Hendriks

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  1. Y Y Ng
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Ng, Y., Baert, M., Pike-Overzet, K. et al. Correction of B-cell development in Btk-deficient mice using lentiviral vectors with codon-optimized human BTK. Leukemia 24, 1617–1630 (2010). https://doi.org/10.1038/leu.2010.140

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