Co-culture of spleen stromal cells with bone marrow mononuclear cells leads to the generation of a novel macrophage subset.

Macrophages adopt diverse activation states depending on the microenvironment. Recently, stromal cells have been demonstrated to be organizers of the microenvironment. Here, using splenic stromal cells to mimic the splenic microenvironment in vivo, we show that spleen stromal cells can programme bone marrow-derived mononuclear cells to differentiate and polarize into a novel macrophage subset. These differentiated macrophages (Diff-Mφ) exhibited pronounced production of IL-10, IL-6 and TNF-α, but diminished the production of IL-12 in response to LPS. The generation of Diff-Mφ depended on cell-cell contact as well as on soluble factors. Diff-Mφ directly suppressed the antigen-non-specific (CD3/CD28) CD4(+) T cell proliferative response and induced cell death of activated CD4(+) T cells. As for cytokine production in CD4(+) T cells, Diff-Mφ promoted IL-10 and IL-17 production, whereas inhibited IL-4 production and did not alter IFN-γ production. Besides, Diff-Mφ also expressed iNOS, CD16/CD32, CD54, CD43, CCR7, CD44, PD-L1 and FasL, which might be involved in the function of Diff-Mφ. These results suggest that splenic microenvironment may physiologically induce a novel type of macrophages differentiation.