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TCRαβ+ double-negative T cells in the kidney are predominantly extravascular and increase in abundance in response to ischemia/reperfusion injury.

TCR+ CD4- CD8- double-negative (DN) T cells are a population of T cells present in low abundance in blood and lymphoid organs, but enriched in various organs including the kidney. Despite burgeoning interest in these cells, studies examining their abundance in the kidney have reported conflicting results. Here we developed a flow cytometry strategy to clearly segregate DN T cells from other immune cells in the mouse kidney and used it to characterise their phenotype and response in renal ischemia-reperfusion injury (IRI). These experiments revealed that in the healthy kidney, most DN T cells are located within the renal parenchyma and exhibit an effector memory phenotype. In response to IRI, the number of renal DN T cells is unaltered after 24 h, but significantly increased by 72 h. This increase is not related to alterations in proliferation or apoptosis. In contrast, adoptive transfer studies indicate that circulating DN T cells undergo preferential recruitment to the post-ischemic kidney. Furthermore, DN T cells show the capacity to upregulate CD8, both in vivo following adoptive transfer and in response to ex vivo activation. Together these findings provide novel insights regarding the phenotype of DN T cells in the kidney including their predominant extravascular location and show that increases in their abundance in the kidney following IRI occur in part due to increased recruitment from the circulation. Furthermore, the observation that DN T cells can upregulate CD8 in vivo has important implications for detection and characterisation of DN T cells in future studies.