The human liver microenvironment shapes the homing and function of CD4+T-cell populations

ObjectiveTissue-resident memory T cells (TRM) are vital immune sentinels that provide protective immunity. While hepatic CD8+TRMhave been well described, little is known about the location, phenotype and function of CD4+TRM.DesignWe used multiparametric flow cytometry, histological assessment and novel human tissue coculture systems to interrogate the ex vivo phenotype, function and generation of the intrahepatic CD4+T-cell compartment. We also used leukocytes isolated from human leukocyte antigen (HLA)-disparate liver allografts to assess long-term retention.ResultsHepatic CD4+T cells were delineated into three distinct populations based on CD69 expression: CD69−, CD69INTand CD69HI. CD69HICD4+cells were identified as tissue-resident CD4+T cells on the basis of their exclusion from the circulation, phenotypical profile (CXCR6+CD49a+S1PR1−PD-1+) and long-term persistence within the pool of donor-derived leukcoocytes in HLA-disparate liver allografts. CD69HICD4+T cells produced robust type 1 polyfunctional cytokine responses on stimulation. Conversely, CD69INTCD4+T cells represented a more heterogenous population containing cells with a more activated phenotype, a distinct chemokine receptor profile (CX3CR1+CXCR3+CXCR1+) and a bias towards interleukin-4 production. While CD69INTCD4+T cells could be found in the circulation and lymph nodes, these cells also formed part of the long-term resident pool, persisting in HLA-mismatched allografts. Notably, frequencies of CD69INTCD4+T cells correlated with necroinflammatory scores in chronic hepatitis B infection. Finally, we demonstrated that interaction with hepatic epithelia was sufficient to generate CD69INTCD4+T cells, while additional signals from the liver microenvironment were required to generate liver-resident CD69HICD4+T cells.ConclusionsHigh and intermediate CD69 expressions mark human hepatic CD4+TRMand a novel functionally distinct recirculating population, respectively, both shaped by the liver microenvironment to achieve diverse immunosurveillance.