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Generation of stable monoclonal antibody–producing B cell receptor–positive human memory B cells by genetic programming

Nature Medicine volume 16, pages 123128 (2010) | Download Citation

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  • A Corrigendum to this article was published on 06 December 2016

This article has been updated

Abstract

The B cell lymphoma-6 (Bcl-6) and Bcl-xL proteins are expressed in germinal center B cells and enable them to endure the proliferative and mutagenic environment of the germinal center. By introducing these genes into peripheral blood memory B cells and culturing these cells with two factors produced by follicular helper T cells, CD40 ligand (CD40L) and interleukin-21 (IL-21), we convert them to highly proliferating, cell surface B cell receptor (BCR)–positive, immunoglobulin-secreting B cells with features of germinal center B cells, including expression of activation-induced cytidine deaminase (AID). We generated cloned lines of B cells specific for respiratory syncytial virus and used these cells as a source of antibodies that effectively neutralized this virus in vivo. This method provides a new tool to study B cell biology and signal transduction through antigen-specific B cell receptors and for the rapid generation of high-affinity human monoclonal antibodies.

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Change history

  • 18 August 2016

    In the version of this article initially published, the article did not mention some restrictions on the availability of reagents. Please note that the retroviral vectors containing BCL-6 and BCL-xL have been generated by a for-profit company, AIMM Therapeutics, which makes the plasmids available. Obtaining the plasmids requires an MTA (http://www.aimmtherapeutics.com/partnering/academic-collaboration/) that includes financial obligations.

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Acknowledgements

We thank B. Hooijbrink for his excellent help with FACS sorting and maintenance of the flow cytometry facility, J. Boes for her excellent work on virus titrations at the Netherlands Vaccine Institute and R. Molenkamp and D. Pajkrt of the Department of Clinical Virology of the Academic Medical Center for helpful discussions. Human cDNA encoding Bcl-xL was kindly provided by S. Korsmeyer (Howard Hughes Medical Institute, Harvard Medical School). Id-3 was a gift from C. Murre (University of California, San Diego). CD40L-L cells were from J. Banchereau (Baylor University). S.A.D. is supported by US National Institutes of Health–National Institute of Allergy and Infectious Diseases grant F32AI063846. M.V.L. is supported by grant OZF-02-004 from the Wilhelmina Research Fund.

Author information

Author notes

    • Sean A Diehl
    •  & Ferenc A Scheeren

    Current addresses: Department of Medicine, University of Vermont, Burlington, Vermont, USA (S.A.D.) and Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Palo Alto, California, USA (F.A.S.).

    • Mark J Kwakkenbos
    •  & Sean A Diehl

    These authors contributed equally to this work.

Affiliations

  1. AIMM Therapeutics, Academic Medical Center, Amsterdam, The Netherlands.

    • Mark J Kwakkenbos
    • , Etsuko Yasuda
    • , Arjen Q Bakker
    • , Caroline M M van Geelen
    • , Hergen Spits
    •  & Tim Beaumont
  2. Department of Cell Biology and Histology, Academic Medical Center, Amsterdam, The Netherlands.

    • Mark J Kwakkenbos
    • , Sean A Diehl
    • , Etsuko Yasuda
    • , Arjen Q Bakker
    • , Caroline M M van Geelen
    • , Ferenc A Scheeren
    • , Hergen Spits
    •  & Tim Beaumont
  3. Department of Pediatrics, The Wilhelmina Children's Hospital, University Medical Center, Utrecht, The Netherlands.

    • Michaël V Lukens
    •  & Grada M van Bleek
  4. Netherlands Vaccine Institute, Bilthoven, The Netherlands.

    • Myra N Widjojoatmodjo
  5. Department of Virology, Biomedical Primate Research Centre, Rijswijk, The Netherlands.

    • Willy M J M Bogers
  6. German Rheumatism Research Center, Berlin, Germany.

    • Henrik Mei
    •  & Andreas Radbruch

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Contributions

M.J.K., S.A.D. and T.B. performed experiments, analyzed data and wrote the paper. H.S. and T.B. organized the research and wrote the paper. E.Y., A.Q.B. and C.M.M.v.G. performed experiments. M.V.L., G.M.v.B., H.M., A.R. and W.M.J.M.B. contributed valuable reagents and developed assays. M.N.W. performed cotton rat experiments. F.A.S. analyzed data and provided valuable suggestions.

Competing interests

S.A.D., F.A.S., E.Y., M.J.K., H.S. and T.B. are listed on patent applications (international patent PCT/NL2008/050333 entitled 'RSV specific binding molecules and means for producing them' and PCT/NL2005/000848 'Means and methods for influencing the stability of cells' (to the patent offices of Japan, the US, Brazil, Canada, Europe, Australia and New Zealand) describing the RSV-specific antibodies and describing the B cell immortalization technology using Bcl-6 and Bcl-xL technology. M.J.K., E.Y., A.Q.B., C.M.M.v.G., T.B. and H.S. are employees of AIMM Therapeutics. H.S. has personal financial interests in AIMM Therapeutics.

Corresponding author

Correspondence to Tim Beaumont.

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DOI

https://doi.org/10.1038/nm.2071