Details

Autor(en) / Beteiligte

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

Titel

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

Ist Teil von

• PloS one, 2010-10, Vol.5 (10), p.e13137

Ort / Verlag

United States: Public Library of Science

Erscheinungsjahr

2010

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• 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.