title: Toll-like receptor ligation for the induction of regulatory B cells.
abstract: Toll-like receptors (TLRs) are key components for the recognition of microorganisms, for the initiation of innate immunity, and for promoting adaptive immune responses. TLR signaling in B cells, in addition to B cell receptor or CD40 ligation, plays an important role in B cell differentiation and activation. In contrast, various infectious agents and/or TLR ligands can also prime B cells to induce tolerance and downregulate inflammatory reactions; those B cells are called regulatory B (Breg) cells and are characterized by a dominant IL-10 production. Several studies have suggested that Breg cells are impaired in patients with autoimmune diseases and allergic asthma. However, the role for TLR ligands in the induction of Breg cells as a potential therapy for some of these inflammatory diseases has not yet been investigated. Here, we provide detailed instructions on how to analyze and validate cytokine production in human and mouse B cells in response to various TLR ligands. Furthermore, we describe an assay to investigate the suppressive properties of TLR-induced B cells to confirm their regulatory B cell status.
authors.affiliation: Department of Parasitology, Cellular Immunology of Helminths, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands.
authors.forename: Luciën E P M
authors.initials: LE
authors.lastname: van der Vlugt
authors.forename: Simone
authors.initials: S
authors.lastname: Haeberlein
authors.forename: Wouter
authors.initials: W
authors.lastname: de Graaf
authors.forename: Tyrill E D
authors.initials: TE
authors.lastname: Martha
authors.forename: Hermelijn H
authors.initials: HH
authors.lastname: Smits
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000032:analysis / Q000276:immunology; D060151:B-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology*; D002469:Cell Separation / Q000379:methods; D018920:Coculture Techniques / Q000379:methods; D016207:Cytokines / Q000032:analysis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D051858:Forkhead Transcription Factors / Q000032:analysis / Q000276:immunology; D006801:Humans; D007113:Immunity, Innate; D016753:Interleukin-10 / Q000032:analysis / Q000276:immunology; D051379:Mice; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology; D051193:Toll-Like Receptors / Q000032:analysis / Q000276:immunology*
chemical_list: D018941:Antigens, CD19; D016207:Cytokines; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D051193:Toll-Like Receptors; D016753:Interleukin-10