- Cai, Curtis;
- Gao, Yu;
- Adamo, Sarah;
- Rivera-Ballesteros, Olga;
- Hansson, Lotta;
- Österborg, Anders;
- Bergman, Peter;
- Sandberg, Johan;
- Ljunggren, Hans-Gustaf;
- Björkström, Niklas;
- Strålin, Kristoffer;
- Llewellyn-Lacey, Sian;
- Price, David;
- Qin, Chuan;
- Grifoni, Alba;
- Wherry, E;
- Sette, Alessandro;
- Aleman, Soo;
- Buggert, Marcus;
- Weiskopf, Daniela
T cells are critical for immune protection against severe COVID-19, but it has remained unclear whether repeated exposure to SARS-CoV-2 antigens delivered in the context of vaccination fuels T cell exhaustion or reshapes T cell functionality. Here, we sampled convalescent donors with a history of mild or severe COVID-19 before and after SARS-CoV-2 vaccination to profile the functional spectrum of hybrid T cell immunity. Using combined single-cell technologies and high-dimensional flow cytometry, we found that the frequencies and functional capabilities of spike-specific CD4+ and CD8+ T cells in previously infected individuals were enhanced by vaccination, despite concomitant increases in the expression of inhibitory receptors such as PD-1 and TIM3. In contrast, CD4+ and CD8+ T cells targeting non-spike proteins remained functionally static and waned over time, and only minimal effects were observed in healthy vaccinated donors experiencing breakthrough infections with SARS-CoV-2. Moreover, hybrid immunity was characterized by elevated expression of IFN-γ, which was linked with clonotype specificity in the CD8+ T cell lineage. Collectively, these findings identify a molecular hallmark of hybrid immunity and suggest that vaccination after infection is associated with cumulative immunological benefits over time, potentially conferring enhanced protection against subsequent episodes of COVID-19.