NK cell function triggered by multiple activating receptors is negatively regulated by glycogen synthase kinase-3 beta

Abstract

Activation of NK cells is triggered by combined signals from multiple activating receptors that belong to different families. Several NK cell activating receptors have been identified, but their role in the regulation of effector functions is primarily understood in the context of their individual engagement. Therefore, little is known about the signaling pathways broadly implicated by the multiple NK cell activation cues. Here we provide evidence pointing to glycogen synthase kinase (GSK)-3 beta as a negative regulator of multiple NK cell activating signals. Using an activation model that combines NKG2D and 2B4 and tests different signaling molecules, we found that GSK-3 undergoes inhibitory phosphorylation at regulatory serine residues by the engagement of NKG2D and 2B4, either individually or in combination. The extent of such phosphorylation was closely correlated with the degree of NK cell activation. NK cell functions, such as cytokine production and cytotoxicity, were consistently enhanced by the knockdown of GSK-3 beta or its inhibition with different pharmacological inhibitors, whereas inhibition of the GSK-3 alpha isoform had no effect. In addition, NK cell function was augmented by the overexpression of a catalytically inactive form of GSK-3 beta. Importantly, the regulation of NK cell function by GSM-3 beta was common to diverse activating receptors that signal through both ITAM and non-ITAM pathways. Thus, our results suggest that GSK-3 beta negatively regulates NK cell activation and that modulation of GSK-3 beta function could be used to enhance NK cell activation. (C) 2015 Elsevier Inc. All rights reserved.