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

• Published in: PloS one 2010-10, Vol.5 (10), p.e13137
• Main Authors: 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
• Format: Article
• Language: English
• Subjects: Analysis; Animals; Antibodies, Monoclonal - biosynthesis; Antigens; B cells; B-Lymphocytes - immunology; Bcl protein; Bcl-6 protein; Bcl-x protein; Biological products; Biological properties; Biological samples; Biology; Biotechnology/Bioengineering; CD19 antigen; CD27 antigen; CD34 antigen; CD38 antigen; CD40 antigen; Cell immortalization; Cell Line, Transformed; Cell Separation; Cells (biology); Cytokines; Endocrinology; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Gastroenterology; Government agencies; Health aspects; Hematopoietic stem cells; Hepatitis; Hepatitis B; Hepatitis B surface antigen; Hepatology; Histology; Immortalization; Immune System; Immunization; Immunoglobulin M; Immunoglobulin M - immunology; Immunoglobulins; Immunology; Immunology/Immune Response; Infections; Infectious diseases; Interleukin 2 receptors; Interleukin 21; Lymphocytes; Lymphocytes B; Lymphoma; Mice; Mice, Inbred BALB C; Monoclonal antibodies; Rodents; Stem cells; T cell receptors; Tetanus; Transplantation; Vaccination; Vaccines
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0013137

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.