Generation of stable monoclonal antibody-producing B cell receptor-positive human memory B cells by genetic programming.
The B cell lymphoma-6 (Bcl-6) and Bcl-xL proteins are expressed in germinal center B cells and enable them to endure the proliferative and mutagenic environment of the germinal center. By introducing these genes into peripheral blood memory B cells and culturing these cells with two factors produced by follicular helper T cells, CD40 ligand (CD40L) and interleukin-21 (IL-21), we convert them to highly proliferating, cell surface B cell receptor (BCR)-positive, immunoglobulin-secreting B cells with features of germinal center B cells, including expression of activation-induced cytidine deaminase (AID). We generated cloned lines of B cells specific for respiratory syncytial virus and used these cells as a source of antibodies that effectively neutralized this virus in vivo. This method provides a new tool to study B cell biology and signal transduction through antigen-specific B cell receptors and for the rapid generation of high-affinity human monoclonal antibodies.
Kwakkenbos
Mark J
MJ
AIMM Therapeutics, Academic Medical Center, Amsterdam, The Netherlands.
Diehl
Sean A
SA
Yasuda
Etsuko
E
Bakker
Arjen Q
AQ
van Geelen
Caroline M M
CM
Lukens
Michaël V
MV
van Bleek
Grada M
GM
Widjojoatmodjo
Myra N
MN
Bogers
Willy M J M
WM
Mei
Henrik
H
Radbruch
Andreas
A
Scheeren
Ferenc A
FA
Spits
Hergen
H
Beaumont
Tim
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D016175:B-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D003564:Cytidine Deaminase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D004789:Enzyme Activation; D005434:Flow Cytometry; D006801:Humans; D007156:Immunologic Memory; D010641:Phenotype; D051560:Proto-Oncogene Proteins c-bcl-6; D051381:Rats; D011947:Receptors, Antigen, B-Cell / Q000096:biosynthesis* / Q000235:genetics; D018357:Respiratory Syncytial Virus Infections / Q000276:immunology; D012136:Respiratory Syncytial Viruses / Q000276:immunology; D013744:Tetanus Toxin / Q000276:immunology; D014161:Transduction, Genetic; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000276:immunology; D051020:bcl-X Protein / Q000096:biosynthesis / Q000235:genetics
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; C494811:BCL2L1 protein, human; C084103:BCL6 protein, human; D004268:DNA-Binding Proteins; D051560:Proto-Oncogene Proteins c-bcl-6; D011947:Receptors, Antigen, B-Cell; D013744:Tetanus Toxin; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; D051020:bcl-X Protein; D003564:Cytidine Deaminase