Effect of Vnut depletion on gene expression during CD4+ T cell differentiation into Th1 cells

Extracellular ATP (eATP), a well-recognized danger signal and immune activator, is implicated in the activation, differentiation and function of T cells, directly or indirectly. Yet, how T cells release ATP themselves and its effects remain poorly investigated. Here we found vesicular nucleotide transporter (VNUT)—critical for vesicular ATP storage/release—is highly expressed in Th1 cells and selectively restricts their differentiation and effector functions. Mechanistically, VNUT facilitates lysosomal ATP import and its extracellular release upon T cell receptor engagement. This eATP then activates the purinergic receptor P2X7R and downstream SRC kinase, triggering a signaling cascade involving heightened Ca2+ influx and hyper-phosphorylation of JNK & FOXO3a, which ultimately impairs Eomes-directed IFN-γ production in Th1 cells. Genetic/pharmacological of VNUT inhibition significantly potentiates Th1 effector functions against Listeria infection and transplanted tumors. These findings identify VNUT as a critical checkpoint in limiting Th1 immunity, coupling vesicular ATP transport to transcriptional control via the P2X7R-JNK-FOXO3a-Eomes axis, offering a target for treating infection and cancer.