Phenotypic and functional characteristics of BMDCs primed under different conditions
We defined the typical phenotypes and characteristics of BMDCs based on two features: the expression of specific costimulatory molecules and the secretion of certain cytokines. As shown in the results, BM cells were triple negative for CD11c, CD80 and MHCII, while BMDCs were triple positive for those markers (Fig. 1B). Interestingly, the expression levels of CD80 and MHCII were lower on DC10 and DClps than DCia (Fig. 2B).
In addition to surface molecules expression, the secretion of certain inflammatory cytokines (IL-10, TGF-β, and IFN-γ) is considered essential for characterizing the tolerogenic potential of BMDCs. BMDCs expressed CCR1, CCR2, and CCR3, all of which bind MCP-319. Thus, we collected BMDC supernatant and examined IL-10, TGF-β, MCP-3 and IFN-γ. Both DC10 and DClps secreted high levels of IL-10 and TGF-β, yet the secretory capacity of these cytokines was markedly lower in DCia. No distinct differences were found in the secretion of MCP-3 or IFN-γ among cells. (Fig. 2A).