Use of Inhibitory Compounds to Dissect the Molecular Pathways Involved in Regulatory B-Cell Differentiation.

Regulatory B cells (Bregs) are IL-10-producing lymphocytes able to suppress inflammatory and autoimmune responses. Pharmacological inhibition of key enzymes within metabolic or signaling pathways enables the identification of factors involved in the differentiation and function of Bregs . Isolation and treatment of splenic B cells derived from IL-10 reporter mice allow fast screening for modulatory compounds influencing IL-10 secretion via flow cytometry. In this chapter, we outline the protocol for the induction of highly potent and metabolically active Bregs using the short-chain fatty acid pentanoate. Moreover, we show how the utilization of inhibitory compounds facilitates the dissection of the engaged pathways in Bregs .