PROLIFERATION AND ACTIVATION OF HUMAN MONONUCLEAR-CELLS INDUCED BY IONOMYCIN IN A SERUM-FREE MEDIUM

Ionomycin was capable of inducing the proliferation and activation of human peripheral blood mononuclear cells (MNC) in serum-free culture medium. Optimal responses, evaluated by tritiated thymidine ([H-3]Thy) incorporation and by HLA-DR antigen expression, were obtained with 0.5-mu-g/ml ionomycin in 72-hour cultures. The two ionomycin-induced effects required the presence of monocytes from the start of the culture. It was observed that no cell adhered to the support after three days of culture in the presence of ionomycin, when compared with an ionomycin-free control. The cells stimulated with ionomycin had a large cytoplasm, were metabolically highly active and had a morphology comparable to that of PHA-stimulated cells. At the end of culture, 23 +/- 4% of cells were DR+; this percentage was only partially found (15 +/- 3%) in all the cell populations studied (T and B lymphocytes, NK cells, monocytes). This led us to believe that this expression occurred in other, probably non-differentiated cells. These later were not stimulated to mature by ionomycin. Thus, ionomycin constitutes an essential early signal, but is insufficient to trigger the complete maturation of cells.