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Immortalization of Human B Cells: Analysis of B Cell Repertoire and Production of Human Monoclonal Antibodies

  • Elisabetta TraggiaiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 901)

Abstract

One of the major challenges in human B cell immunology field has been the objective to establish stable monoclonal cells lines that express the B cell receptor (BCR) on their cell surface and secrete antibodies. Such a system is extremely attractive not only for studying various aspect of BCR signaling but also for the generation of human monoclonal antibody and the analysis of the human B cell repertoire. This chapter describes an efficient method to immortalize and clone human B cells by Epstein–Barr Virus (EBV) transformation.

Key words

B cells Cloning Epstein–Barr Virus Human B cell subsets Human monoclonal antibodies Immortalization Polyclonal stimulation 

References

  1. 1.
    DiLillo DJ, Horikawa M, Tedder TF (2011) B-lymphocytes effector functions in health and disease. Immunol Res 49:281–292PubMedCrossRefGoogle Scholar
  2. 2.
    Jourdan M, Caraux A, Caron G, et al (2011) Characterization of a transitional preplasmablast population in the process of human B cell to plasma cell differentiation. J Immunol 87:3931–3941Google Scholar
  3. 3.
    Kwakkenbos MJ, Diehl SA, Yasuda E, Bakker AQ, van Geelen CMM, Lukens MV, van Bleek GM, Widjojoatmodjo MN, Bogers WMJ, Mei H, Radbruch A, Scheeren FA, Spits H, Beaumont T (2010) Generation of stable monoclonal antibody-producing B cell receptor-positive human memory B cells by genetic programming. Nat Med 16:123–129PubMedCrossRefGoogle Scholar
  4. 4.
    Tiller T, Meffre E, Yurasov S, Tsuiji M, Nussenzweig MC, Wardemann H (2008) Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. J Immunol Methods 329(1–2):112–124PubMedCrossRefGoogle Scholar
  5. 5.
    Meffre E, Wardemann H (2008) B-cell tolerance checkpoints in health and autoimmunity. Curr Opin Immunol 20:632–638PubMedCrossRefGoogle Scholar
  6. 6.
    Steinitz M, Klein G, Koskimies S, Makel O (1977) EB virus-induced B lymphocyte cell lines producing specific antibody. Nature 269: 420–422PubMedCrossRefGoogle Scholar
  7. 7.
    Kozbor D, Roder JC (1981) Requirements for the establishment of high-titered human monoclonal antibodies against tetanus toxoid using the Epstein-Barr virus technique. J Immunol 127:1275–1280PubMedGoogle Scholar
  8. 8.
    McCafferty J, Griffiths AD, Winter G, Chiswell DJ (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554PubMedCrossRefGoogle Scholar
  9. 9.
    Burton DR, Barbas CF III (1994) Human antibodies from combinatorial libraries. Adv Immunol 57:191–280PubMedCrossRefGoogle Scholar
  10. 10.
    Jones PT, Dear PH, Foote J, Neuberger MS, Winter G (1986) Replacing the complementarity-determining regions in a human antibody with those from a mouse. Nature 321:522–525PubMedCrossRefGoogle Scholar
  11. 11.
    Traggiai E, Becker S, Subbarao K, Kolesnikova L, Uematsu Y, Gismondo MR, Murphy BR, Rappuoli R, Lanzavecchia A (2004) An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus. Nat Med 10:871–875PubMedCrossRefGoogle Scholar
  12. 12.
    Traggiai E, Lunardi C, Bason C, Dolcino M, Tinazzi E, Corrocher R, Puccetti A (2010) Generation of anti-NAG-2 mAb from patients’ memory B cells: implications for a novel therapeutic strategy in systemic sclerosis. Int Immunol 22(5):367–374PubMedCrossRefGoogle Scholar
  13. 13.
    Lanzavecchia A, Bernasconi N, Traggiai E, Ruprecht CR, Corti D, Sallusto F (2006) Understanding and making use of human memory B cells. Immunol Rev 211:303–309PubMedCrossRefGoogle Scholar
  14. 14.
    Fingeroth JD, Weis JJ, Tedder TF, Strominger JL, Biro PA et al (1984) Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2. Proc Natl Acad Sci U S A 81: 4510–4514PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Translational ScienceNovartis Institute for Biomedical ResearchBaselSwitzerland

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