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

Autor: Maria Zimmermann, Natalie Rose, John M. Lindner, Hyein Kim, Ana Rita Gonçalves, Ilaria Callegari, Mohammedyaseen Syedbasha, Lukas Kaufmann, Adrian Egli, Raija L. P. Lindberg, Ludwig Kappos, Elisabetta Traggiai, Nicholas S. R. Sanderson, Tobias Derfuss
Rok vydání: 2019
Předmět:
0301 basic medicine
Hemagglutinin Glycoproteins
Influenza Virus
Cell Separation
Lymphocyte Activation
Autoantigens
Cell membrane
Mice
0302 clinical medicine
Methods
Immunology and Allergy
Receptors
Cholinergic
ddc:616
CD20
B-Lymphocytes
biology
Autoantigens/immunology/isolation & purification
Chemistry
autoimmunity
B-Lymphocytes/immunology
Middle Aged
Hemagglutinin Glycoproteins
Influenza Virus/immunology
3. Good health
medicine.anatomical_structure
human monoclonal antibodies
Antigens
Surface
Epitopes
B-Lymphocyte
Antibody
Cell Separation/methods
B-Lymphocyte Subsets/immunology
lcsh:Immunologic diseases. Allergy
Adult
medicine.drug_class
Immunology
B-Lymphocyte Subsets
Hemagglutinin (influenza)
Epitopes
B-Lymphocyte/immunology
Monoclonal antibody
Immunophenotyping
03 medical and health sciences
Antigen
Cell Line
Tumor
Myasthenia Gravis
Antigens
Surface/immunology/isolation & purification
medicine
Animals
Humans
trogocytosis
B cell
Aged
Receptors
Cholinergic/immunology
Myasthenia Gravis/immunology
Viral membrane
Molecular biology
membrane protein antigens
Clone Cells
Mice
Inbred C57BL
030104 developmental biology
biology.protein
lcsh:RC581-607
030215 immunology
Zdroj: Frontiers in Immunology, Vol. 10 (2019) P. 829
Frontiers in Immunology
Frontiers in Immunology, Vol 10 (2019)
ISSN: 1664-3224
DOI: 10.3389/fimmu.2019.00829
Popis: 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. Hemagglutinin-specific B cells isolated this way from vaccinated human donors expressed elevated CD20, CD27, CD71, and CD11c, and reduced CD21, and their secreted antibodies blocked hemagglutination and neutralized viral infection. Antibodies cloned from AChR-capturing B cells derived from patients with myasthenia gravis bound specifically to the receptor on cell membrane. The approach is sensitive enough to detect antigen-specific B cells at steady state, and can be adapted for any membrane antigen.
Databáze: OpenAIRE
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