The immune responses are dysregulated during sepsis. This study aimed to investigate the role of CD57+γδ T cells in the immune pathogenesis of patients with sepsis. We characterized the transcriptomic profile of peripheral γδ T cells from patients with sepsis using smart RNA sequencing. Additionally, we assessed the ratios and immune functional signatures of CD57+γδ T cells in sepsis through flow cytometry. The cytotoxic capacity of the CD57+γδ T-cell subset was also validated in vitro. Our findings revealed a reduction in the proportion of peripheral CD57+γδ T cells in patients with sepsis, which was correlated with clinical outcomes. Compared with CD57-γδ T cells, CD57+γδ T cells exhibited a markedly altered immunophenotype, including decreased expression of costimulatory molecules and increased expression of proinflammatory factors and cytotoxic granules. Our study proposed a potential role of CD57 in sepsis and its utility as a novel biomarker for patients with sepsis.