Lee, J.; Srinivasan, K.; Llinas del Torrent, C.; Ghose, D.; Shah, D. H.; Oliva Hernandez, L.; Wasinger, S.; Chang, M. J.; Yu, W.; Tonai, Y.; Peterson, F. C.; Szpakowska, M.; Chevigne, A.; Volkman, B. F.; Manglik, A.; Lim, W. A.

Chemokines orchestrate the migration of immune cells throughout the body, but harnessing chemokine signaling to engineer targeted cell trafficking is challenging because of their overlapping interactions and broad expression patterns. Here we used structure-guided protein design to engineer orthogonal chemokine-receptor pairs with minimal crosstalk to native systems. T cells engineered with an ortho-receptor selectively infiltrated tissues producing the ortho-chemokine, in vitro and in vivo. By engineering T cells that both sense and secrete the ortho-chemokine, we generated collective swarming behavior in which pioneer cells attract follower cells via an ortho-chemokine gradient self-organized by positive feedback. By providing orthogonal genetic control over both the generation and sensing of chemokine gradients, this system enables the construction of autonomous spatial targeting programs in engineered cells.