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This second edition provides new and updated chapters useful for the study of Regulatory B cells. Organized in four sections, chapters detail basic methods for the isolation and immunophenotypical analysis of these cells, experimental approaches for the ex vivo generation/expansion of IL-10 producing B cells, molecular biology techniques for the analysis of IL-10 expression and production, and animal models mimicking pathologic settings. Written in the highly successful Methods in Molecular Biology series format, chapters include an introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Regulatory B Cells: Methods and Protocols, Second Edition aims to be useful to the scientific community and serve toclarify some unsolved aspects of Regulatory B Cells research.
List of contents
Purification and characterization of murine MZ and T2-MZP cells.- Purification and immune-phenotyping of B-1 cells from body cavities of mice.- Purification and immunophenotypic characterization of murine plasma cells.- Purification of murine and human IL-10 producing B cells from different anatomical compartments.- Purification and immunophenotypic characterization of human CD19+CD24hiCD38hi and CD19+CD24hiCD27+ B cells.- Detection of IL-10 in murine B cells: in vitro and in vivo techniques.- Detection and quantification of transforming growth factor-beta1 produced by murine B cells: pros and cons of different techniques.- IL-35 detection in B cells at the mRNA and protein level.- Characterization and activation of Fas Ligand producing mouse B cells and their killer exosomes.- Characterization and activity of TIM-1 and IL-10-reporter expressing regulatory B cells.- New method for the expansion of highly purified human regulatory granzyme B-expressing B cells.- Characterization of the cell surface phenotype and regulatory activity of B cell IgD low (BDL) regulatory B cells.- Use of Toll-like receptor (TLR) ligation to characterize human regulatory B cells subsets.- Use of co-cultures for the study of cellular interactions influencing B cell regulatory functions.- Use of inhibitory compounds to dissect the molecular pathways involved in regulatory B cell differentiation.- Methods to study the transcriptome of regulatory B cells.- Purification of murine IL-10+ B cells for analyses of biological functions and transcriptomics.- il-10 gene locus DNA methylation in regulatory B cells.- B cell commitment to IL-10 production: the Vert-X Il10egfp mouse.- Regulatory B cells in experimental mouse models of arthritis.- Regulatory and IgE+ B cells in allergic asthma.- Regulatory B cells in type 1 diabetes.- Analysis of regulatory B cells in experimental autoimmune uveitis.- Purification and immunophenotypic characterization of human CD24hiCD38hi and CD24hiCD27+ regulatory B cells in transplantation.
