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

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

PloS one (2010-10-20)
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, James P Di Santo, Tim Beaumont, Carlos A Guzman, Hergen Spits
ABSTRACT

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 time-consuming 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. 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 BCL-XL 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. 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.

MATERIALS
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Brand
Product Description

Sigma-Aldrich
Interleukin-21 from mouse, ≥97% (SDS-PAGE), recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture