Use of Cocultures for the Study of Cellular Interactions Influencing B-Cell Regulatory Functions.

Although IL-10-producing B cells have been shown to play key roles in regulating immune responses involved in autoimmunity, inflammation, and cancer, the mechanisms at the base of the generation and maintenance of the pool of regulatory B cells are still poorly characterized. Several evidences show that the cross talk between B cells and other immune cell types promotes IL-10 production by B lymphocytes. Soluble mediators released into the microenvironment, together with direct cell-cell contact, are key signals in the process of regulatory B-cell development and differentiation. Here we describe the methods required to follow IL-10-producing B cells in MC- and MDSC-B-cell cocultures as examples of in vitro systems that induce the expansion of the regulatory B-cell population. These protocols can be also adapted for the study of other immune cell systems.