Description
This volume details in methods to assess memory B cell formation and function in mice and humans. Chapters guide readers through tetramer-based methods to assess antigen-specific memory B cell dynamics in humans and mice in different vaccine, next-generation deep-sequencing, single-cell techniques to assess epigenomic, VDJ landscapes, lymph node aspirates from humans, advanced imaging, murine models to determine memory B cells formation, and bioinformatic techniques and in silico modelling of memory b cell formation. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Memory B-Cells: Methods and Protocols aims to ensure successful results in the further study of this vital field.
Table of Contents
Using ex vivo tonsil organoids to study memory B cells.- Ultrasound Guided Fine Needle Biopsy of Human Axillary Lymph Nodes to assess B cell responses to Vaccination.- Human B cell receptor repertoire sequencing.- Visualising epigenetic modifications and nuclear bodies by immunofluorescence staining in naïve, activated and memory B cell subsets.- Measuring B cell Chromatin Accessibility by One-Step ATAC-seq.- Epigenomic profiling of B cell subsets by CUT&Tag.- In vivo CRISPR/Cas9-mediated gene ablation in murine B cells.- Isolation of rare antigen-specific memory B cells via antigen tetramers.- Detection of lymphocytic choriomeningitis virus-specific memory B cells using antigen tetramers.- B cell receptor transgenic mice as tools to study memory B cells.- ELISpots for detecting antigen-specific memory B cells in Infection or Autoimmunity.- Intracellular flow cytometry (“phosphoflow”) to assess signal transduction in rare populations such as memory B cell subsets and plasma cells.- Quantifying human naïve B cell proliferation kinetics and differentiation in controlled in vitro cell culture.- Flow cytometric identification of human IgE+ B lineage subsets.- Identification of IgG1 and IgG3 Allotypes by PCR and Sanger Sequencing.-Assessing the survival of human plasma cells ex vivo.- Conversion of mouse-derived hybridomas to Tasmanian devil recombinant IgG antibodies.
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