CD13/N-aminopeptidase is involved in the development of dendritic cells and macrophages from cord blood CD34+ cells

Abstract

Expression of CD13/N-aminopeptidase may reflect cell activation and growth. We examined its role regarding cell growth in cultures of cord blood CD34+ cells with stem cell factor/Flt-3 ligand/granulocyte-macrophage colony-stimulating factor/tumor necrosis factor-. Indeed, 82% ± 6% of cells from culture day 5 were CD13hi, 25% ± 8% of which were still Lin−. About 50% of CD13hiLin− cells, which comprise progenitors of dendritic cells (DC), monocytes/macrophages and granulocytes, and 30% of CD13loLin− cells were CD34+. Sorted CD34+CD13hiLin− cells, cultured further for 7 days with the same cytokines, expanded 31-fold and CD34-CD13hiLin− cells 7-fold, but CD34+CD13loLin− and CD34−CD13loLin− cells did not grow. Thus, cell growth correlated with CD13 expression, all the more so that cells were CD34+. Actinonin, the most potent N-aminopeptidase inhibitor, was used to engage CD13 on sorted CD13hiLin− cells and on culture day-7 bulk cells. In both cases, this resulted in reversible cell growth arrest, with 30% to 60% fewer cells in the G2/S-M phase than in controls. Interestingly, similar effects were noted with CD13 monoclonal antibody TÜK1, which does not inhibit N-aminopeptidase activity, but not with N-aminopeptidase-blocking antibodies WM15 and F23. All cycling cells appeared susceptible to actinonin, which induced cell apoptosis at the same time as Bcl-2 was downregulated and caspase-3 activity increased, but finally percentages and yields of DC and macrophage precursors were affected more than those of granulocytic cells. Thus, through engagement of N-aminopeptidase enzymatic site but possibly also of an independent determinant, CD13 plays a role in the growth of DC/macrophage progenitors and precursors.