Potent suppression of natural killer cell response mediated by the ovarian tumor marker CA125

Objectives. CA125 expresses specific oligosaccharides that can inhibit the cytotoxicity of human natural killer (NK) cells. The current study was undertaken to determine the ability of CA125 to modulate NK cell-mediated cytotoxicity. Methods. CA125 was isolated from OVCAR-3 cells and its purity was determined by ELISA and ultra-sensitive mass spectrometric analysis. Peripheral blood-derived NK were treated with CA125 and standard cytotoxicity assays were performed using Cr-51-labeled K562 cells as targets. The expression of cell surface and intracellular markers on NK cells was determined by either flow cytometry or Western blot analysis. Results. NK cells incubated with CA125 for 72 It exhibited a 50-70% decrease in the lysis of K562 targets. Incubation with CA125 for 4 It and 24 h had no effect on NK-mediated cytolysis. Inhibition of NK function was observed at CA125 concentrations (10,000-100,000 U/ml) that are expected to be significantly lower than those observed in the tumor microenvironment. Co-stimulation with IL-2 did not abrogate the NK inhibitory response of CA125. CA125 did not reduce proliferation or induce apoptosis of NK cells and alter the expression of p561ck, phospholipase C-gamma 1, ZAP70, or CD3 zeta. CA125 did, however, induce major downregulation of CD16 and minor decrease in expression of CD94/NKG2A. Conclusions. Our ongoing research and recent work performed by other laboratories highlights the potential physiologic role of this mucin. Based on the data presented here, it is likely that the tumor-derived CA125 acts as a suppressor of the immune response that is directed against the ovarian tumors. (c) 2005 Elsevier Inc. All rights reserved.