Optical Manipulation of T-lymphocyte Activation

My current research seeks to control T-cell activation. Manipulation of the interface between T-cells and antigen presenting cells (APCs), also known as the immunological synapse(IS), is critical to engineering T-cell effector function. An increasing body of evidence suggests that the spatial organization of TCR:pMHC and CD28:CD80, two essential ligand-receptor pairings within the IS, greatly impact T-cell activation. Lck affects both TCR and CD28 signaling, thus the focus of our study. We recapitulated these spatial cues with microscale patterned planar surfaces, utilizing indirect capturing to present pMHC and CD80 to naive T-cells. We described here the incorporation of photoactivable pMHC into the multi-component patterned surface for spatiotemporal control of Lck activation or phosphorylation (pLck). We also presented a FRET reporter for pLck kinase activities. Previous work from our group showed that differential spatial organization of pLck between human and mouse naive T-cells could explain the observed differences in IL-2 secretion. And preliminary data from our group highlighted pronounced differences in Lck diffusion rates via FRAP. My pLck reporter would not only give its diffusion rate but also the state of its kinase activity.