Generation of human antigen-specific monoclonal IgM antibodies using vaccinated "human immune system" mice

Pablo D. Becker, Nicolas Legrand, Caroline M.M. van Geelen, Miriam Noerder, Nicholas D. Huntington, Annick Lim, Etsuko Yasuda, Sean A. Diehl, Ferenc A. Scheeren, Michael Ott, Kees Weijer, Heiner Wedemeyer, Jamesdi P. di Santo, Tim Beaumont, Carlos A. Guzman, Hergen Spits

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Background: Passive transfer of antibodies not only provides immediate short-term protection against disease, but also can be exploited as a therapeutic tool. However, the 'humanization' of murine monoclonal antibodies (mAbs) is a timeconsuming and expensive process that has the inherent drawback of potentially altering antigenic specificity and/or affinity. The immortalization of human B cells represents an alternative for obtaining human mAbs, but relies on the availability of biological samples from vaccinated individuals or convalescent patients. In this work we describe a novel approach to generate fully human mAbs by combining a humanized mouse model with a new B cell immortalization technique. Methodology/Principal Findings: After transplantation with CD34+CD38- human hematopoietic progenitor cells, BALB/c Rag2-/-IL-2Rγc-/- mice acquire a human immune system and harbor B cells with a diverse IgM repertoire. "Human Immune System" mice were then immunized with two commercial vaccine antigens, tetanus toxoid and hepatitis B surface antigen. Sorted human CD19+CD27+ B cells were retrovirally transduced with the human B cell lymphoma (BCL)-6 and BCLXL genes, and subsequently cultured in the presence of CD40-ligand and IL-21. This procedure allows generating stable B cell receptor-positive B cells that secrete immunoglobulins. We recovered stable B cell clones that produced IgM specific for tetanus toxoid and the hepatitis B surface antigen, respectively. Conclusion/Significance: This work provides the proof-of-concept for the usefulness of this novel method based on the immunization of humanized mice for the rapid generation of human mAbs against a wide range of antigens.

Original languageEnglish
Article numbere13137
JournalPLoS ONE
Issue number10
Publication statusPublished - 4 Nov 2010
Externally publishedYes

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