Nanoconfinement of microvilli alters gene expression and boosts T cell activation

Significance Microvilli are used by immune cells to sense the surface features of pathogens and antigen presenting cells. However, microvilli’s contribution in T cell signaling and activation is largely unknown. Here, we introduce a material-based platform for induction of microvilli formation in T cells, in which the dimensions of the microvilli can be controlled by tuning the dimensions of the nanotopographical features, such as pore depth, pore size, and interpore distance. We demonstrate the direct causality between microvilli formation and altered gene expression in T cells. We discover that the size of the microvilli critically influences T cell receptor agonistic independent signaling in T cells. The results provide a physical strategy for T cell activation and expansion for immunotherapy applications. T cells sense and respond to their local environment at the nanoscale by forming small actin-rich protrusions, called microvilli, which play critical roles in signaling and antigen recognition, particularly at the interface with the antigen presenting cells. However, the mechanism by which microvilli contribute to cell signaling and activation is largely unknown. Here, we present a tunable engineered system that promotes microvilli formation and T cell signaling via physical stimuli. We discovered that nanoporous surfaces favored microvilli formation and markedly altered gene expression in T cells and promoted their activation. Mechanistically, confinement of microvilli inside of nanopores leads to size-dependent sorting of membrane-anchored proteins, specifically segregating CD45 phosphatases and T cell receptors (TCR) from the tip of the protrusions when microvilli are confined in 200-nm pores but not in 400-nm pores. Consequently, formation of TCR nanoclustered hotspots within 200-nm pores allows sustained and augmented signaling that prompts T cell activation even in the absence of TCR agonists. The synergistic combination of mechanical and biochemical signals on porous surfaces presents a straightforward strategy to investigate the role of microvilli in T cell signaling as well as to boost T cell activation and expansion for application in the growing field of adoptive immunotherapy.