Monocytes immunoselected via the novel monocyte specific molecule, CD300e, differentiate into active migratory...

Monocytes, immunoselected using MMRI-1, a monoclonal antibody specific for CD300e, were used to generate dendritic cells (DC). These CD300e immunoselected monocyte-derived DC (MoDC) were compared phenotypically and functionally to CD14 immunoselected MoDC. CD300e and CD14 immunoselected mature MoDC expressed similar levels of the DC marker, CD83 and costimulatory molecules, CD80, CD86, and CD40. Both preparations took up soluble antigen with similar efficiency by pinocytosis and receptor mediated uptake. The CD300e and CD14 immunoselected MoDC also induced comparable CD4+ T lymphocyte allogeneic responses and recall responses to tetanus toxoid. Similar magnitude CD8 T lymphocyte responses to the naive antigen, MART-1 and the recall antigen, FMP, were induced by both MoDC preparations. Cytokine secretion by each type of MoDC preparation was similar; each secreted interleukin-12, tumor necrosis factor-alpha, and low levels of interferon-gamma but in most cases no interleukin-10. Migration studies confirmed that both types of MoDC migrated towards the chemokine, CCL21 although CD300e immunoselected showed greater migration. Overall, the CD14 immunoselected MoDC had higher spontaneous background migration, compared with the CD300e immunoselected MoDC. Differential signaling from the antibodies used to immunoselect the monocytes may account for the slight differences in migratory capacity. These data identify the CD300e antigen as another monocyte-specific marker that can be used to purify monocytes for differentiation into functionally active MoDC.