Antigen Extraction and B Cell Activation Enable Identification of Rare Membrane Antigen Specific Human B Cells.

ABSTRACT: Determining antigen specificity is vital for understanding B cell biology and for producing human monoclonal antibodies. We describe here a powerful method for identifying B cells that recognize membrane antigens expressed on cells. The technique depends on two characteristics of the interaction between a B cell and an antigen-expressing cell: antigen-receptor-mediated extraction of antigen from the membrane of the target cell, and B cell activation. We developed the method using influenza hemagglutinin as a model viral membrane antigen, and tested it using acetylcholine receptor (AChR) as a model membrane autoantigen. The technique involves co-culturing B cells with adherent, bioorthogonally labeled cells expressing GFP-tagged antigen, and sorting GFP-capturing, newly activated B cells. H

SUBMITTER: Zimmermann M 

PROVIDER: S-EPMC6477023 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature