Single-cell analysis of the dynamics and functional outcomes of interactions between human natural killer cells and target cells

Abstract

2 Graphical abstractWe used arrays of subnanoliter wells (nanowells) to monitor the dynamics of individual NK celltarget cell interactions and quantify the resulting cytolytic and secretory responses. Insight statementThe cytolytic and secretory activities of natural killer (NK) cells are important components of the innate immune response against potentially harmful target cells. We developed a nanowell-based platform to efficiently acquire multiparametric measurements of encounters between individual NK cells and tumor target cells. The integrated profiles of contact behavior, motility, lysis, and secretion yielded new insights into the heterogeneities and correlations of these parameters at the single-cell level. We found that NK cells lyse targets in an autonomous, history-dependent manner, and that the target cell-induced secretory activity of individual NK cells associated with their migratory characteristics. This technology can be adapted to investigate, with single-cell resolution, the dynamic and functional attributes of other cellular systems of interest.

Classification of human natural killer cells based on migration behavior and cytotoxic response

“…Although NK cell cytotoxicity was directly observable, our experimental setup did not allow measurement of cytokine secretion and, therefore, it was not possible to correlate this to certain types of behavior such as killing or nonkilling. However, a recent study demonstrated that there was no difference in the levels of MIP-1␤ and IFN-␥ production at the single-cell level between lytic and nonlytic interactions, 30 suggesting that no differences in the levels of cytokine secretion would have been observed between lytic and nonlytic NK cells.…”

Longitudinal multiparameter assay of lymphocyte interactions from onset by microfluidic cell pairing and culture

“…made it feasible to resolve the relationships between different immune responses, particularly between cellular motility, cytotoxicity, and secretory activity (9,10,12), two primary limitations inherent in their operation principles have precluded the ability to study early signaling dynamics and their correlation to subsequent functional behaviors. First, generation of cell-cell interactions is governed by stochastic cell loading that limits conjugate formation efficiency and yields low-throughput-per-footprint area, which in turn severely limits the spatiotemporal resolution for dynamic imaging (i.e., number of cells that can be imaged with high temporal resolution).…”

“…A common approach has been to isolate discrete numbers of cells in spatial confinements [such as microwells (9)(10)(11)(12), microchambers (13), or droplets (14)] and monitor their interactions with multiple measurements. Although these approaches have…”

Ultrasound-Induced Cell–Cell Interaction Studies in a Multi-Well Microplate

“…An alternative strategy is to dispense several cells per well and then to study individual interactions between two or more cells. This has been used for studying interaction between natural killer cells and different types and numbers of target cells [9,[11][12][13][14], and also for migration studies [15].…”