title: Use of cocultures for the study of cellular interactions influencing B cell regulatory functions.
abstract: Many immunological processes are contextually controlled by complex interactions among different cell types. Several studies have shown that B cells produce the immune regulatory cytokine IL-10 in response to different external stimuli but also to immune-mediated signals. Endogenous signals that derive from the cross talk between B lymphocytes and other cells of the immune system can affect IL-10 production by B cells in both physiological and pathological conditions. With the aim to provide a guide for the study of how partner cells can induce IL-10-producing B cells, here we describe the protocols to investigate IL-10 production at a single cell level in a dendritic cell-B cell coculture in vitro system. These methods are a proof of concept that can be easily extrapolated and adapted to the study of the interaction between B cells and other immune cell types.
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le M. Kolbe 4, 33100, Udine, Italy, francesca.mion@uniud.it.
authors.forename: Francesca
authors.initials: F
authors.lastname: Mion
authors.forename: Federica
authors.initials: F
authors.lastname: Benvenuti
authors.forename: Carlo
authors.initials: C
authors.lastname: Pucillo
authors.forename: Gaetano
authors.initials: G
authors.lastname: Vitale
mesh_terms: D000818:Animals; D060151:B-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D002450:Cell Communication; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000295:instrumentation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D016753:Interleukin-10 / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction / Q000379:methods; D016707:Tissue Fixation / Q000379:methods; D014158:Transcription, Genetic
chemical_list: D016753:Interleukin-10