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

Distinct gene expression patterns correlate with developmental and functional traits of iNKT subsets

Invariant natural killer T (iNKT) cells comprise a subpopulation of innate lymphocytes developing in thymus. A new model proposes subdividing murine iNKT cells into iNKT1, 2 and 17 cells. Here, we use transcriptome analyses of iNKT1, 2 and 17 subsets isolated from BALB/c and C57BL/6 thymi to identify candidate genes that may affect iNKT cell development, migration or function. We show that Fcɛr1γ is involved in generation of iNKT1 cells and that SerpinB1 modulates frequency of iNKT17 cells. Moreover, a considerable proportion of iNKT17 cells express IL-4 and IL-17 simultaneously. The results presented not only validate the usefulness of the iNKT1/2/17-concept but also provide new insights into iNKT cell biology