Purification and immunophenotypic characterization of human B cells with regulatory functions

Abstract

The analysis of human B cell populations of the blood relies on the expression of surface markers, mainly CD19, CD24, CD38, and CD27. According to these surface markers, three main B cell subsets can be identified in the blood: immature transitional B cells (CD19 + CD24 high CD38 high), naïve B cells (CD19 + CD24 int CD38 int) that have not encountered an antigen, and memory B cells (CD19 + CD27 +). To date, human B cells with regulatory functions have been essentially described within the CD24 high CD38 high transitional B cell subset. CD24 high CD38 high transitional B cells are able to produce interleukin 10 (IL-10) and to regulate in vitro Th1 and Th17 CD4 + T cell activation. Here, we provide the methods to analyze and purify the CD24 high CD38 high transitional B cell subset for further in vitro experiments. We also provide a reliable method to detect B cell IL-10 production using intracellular cytokine staining. © 2014 Springer Science+Business Media New York.