Memory B-Cells

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.

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.