Actin conformation dynamics precede force generation in cytotoxic T lymphocytes
Rochussen, A. M.; Kebabci, A. O.; Winkel, A. K.; Franze, K.; Fletcher, D. A.; Griffiths, G. M.; Lippert, A. H.
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Cytotoxic T lymphocytes (CTLs) are cells of the adaptive immune system that are able to recognise and kill cancerous or virally infected target cells. The biochemical basis of CTL-mediated killing has been examined in great detail. Recent work has underscored the importance of physical forces in T cell function too, but how CTLs sense and exert force during migration and killing is not fully understood. Here, by expressing actin conformation probes based on the CH domain of utrophin, we directly visualize regions of altered F-actin conformation in primary CTLs during migration and killing. By combining these probes with traction force microscopy, we correlate external force with the internal cytoskeleton. We show that actin conformation is regulated upstream of force production, and that force exertion at the cytotoxic immune synapse temporally follows actin conformation dynamics. Our work offers novel insight into the relationship between cellular force production and F-actin conformation in important primary immune cells.
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