Cytokines play critical roles in homeostatic control of health and they are integral for the creation and maintenance of a myriad of disease states. Their ultra-low concentration, often in the picomolar range, and extremely dynamic transient secretion process place stringent demands on cytokine quantification. We developed a nanoparticle-based strategy to detect trace cytokine secretion from individual, single live cells, for which we coined the term “OnCELISA”. Using a capture surface on the cell membrane and fluorescent magnetic nanoparticles as assay reporters, our universal OnCELISA assay achieved the sensitivity 0.1 pg mL-1, an over 10-fold enhancement, compared to state-of-the-art. The sensitive OnCELISA cell labelling made it possible to select and sort different cell types to determine highly cytokine - secreting cell subpopulations . The capture surfaces on cell membranes did not show noticeable effect on cell viability and their subsequent proliferation. The capability to specifically select such highly cytokine-secreting cells and purify their populations is pivotal for their use in multicellular pathologies such as atherosclerosis. Accordingly, we used this new approach to label cytokine secretion from vascular tissues of apolipoprotein E-/- mice; an in vivo model of atherosclerosis. In response to lipopolysaccharide, we observed increased capture of cytokine using this model. With the capacity of monitoring multiple cytokine secretions (IL-6 and IL-1β)), our OnCELISA method is able to probe how the individual cells and tissues secrete cytokines as they respond in real time to the surrounding signals.
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