This volume looks the current advanced protocols used to study aspects of the B cell receptor (BCR). The chapters in this book cover topics such as the mutant of BCR repertoire to understand antibody evolution; interactions between B cells and viruses; mechanical force during BCR activation; B cell signaling using flow cytometry; confocal microscopy, total internal reflection microscopy and intravital two-photon microscopy; and the methods used to study critical cell components related with B cell activation. 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 tips on troubleshooting and avoiding known pitfalls.Cutting-edge and practical, B Cell Receptor Signaling: Methods and Protocols is a valuable resource for everyone in the scientific community currently working in the B cell field.
Preface...Table of Contents...Contributing Authors...1. Activation-Induced Cytidine Deaminase Aided In Vitro Antibody EvolutionLili An, Chuan Chen, Ruiqi Luo, Yun Zhao, and Haiying Hang 2. Analyzing Mouse B-Cell Responses Specific to LCMV InfectionYaxing Hao, Zhirong Li, Yifei Wang, Xiaobing Liu, and Lilin Ye3. Expression of Exogenous Genes in Murine Primary B Cells and B Cell Lines Using Retroviral Vectors Zhiyong Yang and Christopher D. C. Allen 4. Biophysical Techniques to Study B Cell Activation: Single Molecule Imaging and Force Measurements Ivan Rey, David A. Garcia, Brittany A. Wheatley, Wenxia Song, and Arpita Upadhyaya 5. DNA-Based Probes for Measuring Mechanical Forces in Cell-Cell Contacts: Application to B Cell Antigen Extraction from Immune SynapsesKatelyn M. Spillane and Pavel Tolar6. Deriving Quantitative Cell Biological Information from Dye-Dilution Lymphocyte Proliferation ExperimentsKoushik Roy, Maxim Nikolaievich Shokhirev, Simon Mitchell, and Alexander Hoffmann7. Flow Cytometry Analysis of mTOR Signaling in Antigen-Specific B CellsQizhao Huang, Haoqiang Wang, Lifan Xu, Jianjun Hu, Pengcheng Wang, Yiding Li, and Lilin Ye8. Ex Vivo Culture Assay to Measure Human Follicular Helper T (Tfh) Cell-Mediated Human B Cell Proliferation and Differentiation Xin Gao, Lin Lin, and Di Yu ;9. B Cell Receptor Signaling and Compartmentalization by Confocal MicroscopyAnurag R. Mishral and Akanksha Chaturvedi 10. Investigate Early Activation Events of B Cell Receptor Signaling by Membrane Associated Antigen System and Total Internal Reflection Fluorescence MicroscopyXiaoming Bai, Xiaodong Zhao, and Chaohong Liu 11. Imaging the Interactions between B Cells and Antigen-Presenting Cellsal,Jia C. Wang, Madison Bolger-Munro, and Michael R. Gold12. In Vivo Tracking of Particulate Antigen Localization and Recognition by B Lymphocytes at Lymph NodesYolanda R. Carrasco13. Study B Cell Antigen Receptor Nano-scale Organization by In Situ Fab Proximity Ligation AssayKathrin KlaÌ sener, Jianying Yang, and Michael Reth14. Single Particle Tracking of Cell Surface ProteinsLaabiah Wasim and Bebhinn Treanor15. The Use of Intravital Two-Photon and Thick Section Confocal Imaging to Analyze B Lymphocyte Trafficking in Lymph Nodes and SpleenChung Park, Il-Young Hwang, and John H. Kehrl16. Time-lapse Forster Resonance Energy Transfer Imaging by Confocal Laser Scanning Microscopy for Analyzing Dynamic Molecular Interactions in the Plasma Membrane of B CellsHae Won Sohn and Joseph Brzostowski17. Understanding of B Cell Receptor Signaling through a Photo-Activatable Antigen Presentation SystemJing Wang, Zhengpeng Wan, and Wanli Liur18. Use of Streptolysin O-induced Membrane Damage as a Method of Studying the Function of Lipid Rafts during B Cell ActivationHeather Miller and Wenxia Song19. Visualization and Quantitative Analysis of the Actin Cytoskeleton upon B Cell ActivationVid Sustar, Marika Vainio, Pieta K. Mattila