Dendritic cells as the terminal stage of monocyte differentiation

Abstract

Monocytes (MO) cultured for > or =5 days with either macrophage-CSF (M-CSF) or granulocyte macrophage (GM)-CSF and IL-4 differentiated without concomitant proliferation into CD14+ macrophages (Mphi) or CD1a+ dendritic cells (DC), respectively. When adherent and nonadherent CD14high Mphi from M-CSF cultures were separated and cultured further in cytokine-free medium or with GM-CSF/IL-4, most cells from both fractions that were exposed to GM-CSF/IL-4 acquired CD1a expression and DC morphology and function. Conversely, GM-CSF/IL-4 withdrawal at day 5 and additional culture of sorted CD1a+ DC for 2 to 7 days in cytokine-free medium led to cells rapidly becoming adherent CD1a-CD14+ Mphi. Replacing GM-CSF/IL-4 with M-CSF hastened the conversion of DC to Mphi without increasing cell numbers. CD1a+CD14-CD83+ mature DC were induced by a > or =2-day exposure to MO-conditioned medium, LPS, or TNF-alpha/IL-1beta. Upon cytokine removal or culture with M-CSF, DC that had been pushed to maturation by conditioned medium or LPS remained stable or died in the new environment. TNF-alpha/IL-1beta-driven DC displayed heterogeneous CD83 expression and could thus be sorted into CD83high and CD83low/- cells; in cytokine-free medium or in M-CSF, most CD83low/- cells converted to Mphi, whereas most CD83high cells remained nonadherent CD1a+CD14- or died and thus appeared truly terminally differentiated. Hence, MO are precursors of Mphi as well as of DC, with each cell type having the capability to convert into the other until late in the differentiation/maturation process. Accordingly, the cytokine environment and the presence of differentiation and/or other stimulatory signals may be the "final decision-making factors" determining whether these cells will acquire DC or Mphi characteristics and function.