Exhausted CD8⁺ T cells exhibit low and strongly inhibited TCR signaling during chronic LCMV infection

Chronic viral infections are often associated with impaired CD8⁺ T cell function, referred to as exhaustion. Although the molecular and cellular circuits involved in CD8⁺ T cell exhaustion are well defined, with sustained presence of antigen being one important parameter, how much T cell receptor (TCR) signaling is actually ongoing in vivo during established chronic infection is unclear. Here, we characterize the in vivo TCR signaling of virus-specific exhausted CD8⁺ T cells in a mouse model, leveraging TCR signaling reporter mice in combination with transcriptomics. In vivo signaling in exhausted cells is low, in contrast to their in vitro signaling potential, and despite antigen being abundantly present. Both checkpoint blockade and adoptive transfer of naïve target cells increase TCR signaling, demonstrating that engagement of co-inhibitory receptors curtails CD8⁺ T cell signaling and function in vivo.