title: IL-10 detection in murine B cells: pros and cons of the different techniques.
abstract: With the recently increased understanding of the roles of B cells in immune response and autoimmune pathogenesis, various techniques have been used for the detection of IL-10 production in B cells. In this chapter, we describe several commonly used methods for the effective detection of IL-10 in B cells at both mRNA and protein levels, including quantitative PCR analysis, intracellular staining of IL-10 by flow cytometry, ELISA assay for secreted IL-10 detection, and ELISPOT assay of enumerating IL-10-producing B cells. Moreover, we provide a detailed protocol for the detection of IL-10-producing B cells in situ by immunofluorescence microscopy. Together, the application of these described methods for the detection of IL-10 will facilitate the characterization of B cell subsets with regulatory functions and enhance our current understanding on the critical roles of B cells in immune response and autoimmune development.
authors.affiliation: Department of Pathology, Queen Mary Hospital, The University of Hong Kong, Room 234, University Pathology Building, Pokfulam Road, Hong Kong, Hong Kong.
authors.forename: Xiang
authors.initials: X
authors.lastname: Lin
authors.forename: Jun
authors.initials: J
authors.lastname: Deng
authors.forename: Liwei
authors.initials: L
authors.lastname: Lu
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D016753:Interleukin-10 / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: D012333:RNA, Messenger; D016753:Interleukin-10