NK cell activation is regulated by a balance between activating and inhibitory signals. To address the question of how these signals are spatially integrated, we created a computer simulation of activating and inhibitory NK cell immunological synapse (NKIS) assembly, implementing either a “quantity-based” inhibition model or a “distance-based” inhibition model. The simulations mimicked the observed molecule distributions in inhibitory and activating NKIS and yielded several new insights. First, the total signal is highly influenced by activating complex dissociation rates but not by adhesion and inhibitory complex dissociation rates. Second, concerted motion of receptors in clusters significantly accelerates NKIS maturation. Third, when the potential of a cis interaction between Ly49 receptors and MHC class I on murine NK cells was added to the model, the integrated signal as a function of receptor and ligand numbers was only slightly increased, at least up to the level of 50% cis-bound Ly49 receptors reached in the model. Fourth, and perhaps most importantly, the integrated signal behavior obtained when using the distance-based inhibition signal model was closer to the experimentally observed behavior, with an inhibition radius of the order 3–10 molecules. Microscopy to visualize Vav activation in NK cells on micropatterned surfaces of activating and inhibitory strips revealed that Vav is only locally activated where activating receptors are ligated within a single NK cell contact. Taken together, these data are consistent with a model in which inhibitory receptors act locally; that is, that every bound inhibitory receptor acts on activating receptors within a certain radius around it.

Proper functioning of the immune system depends on coordinated intercellular communication that occurs in the "immunological synapse". This is the contact area of cell-cell conjugates where information is transferred via segregated clusters of proteins. Vast experimental datasets on its assembly are available, and mathematical and computational modeling represent a useful tool to integrate all this information. We created a stochastic computer simulation to study immunological synapse formation. Using this simulation we show that the initial locations of adhesion molecules and T cell receptors (TCRs) at the moment of contact influence the inversion of the immature synapse into a mature synapse, and that movement of large clusters of TCRs towards the center of the contact area accelerates this inversion.