In mice, the ability of naive T (TN) cells to mount an effector response correlates with TCR sensitivity for self-derived Ags, which can be quantified indirectly by measuring surface expression levels of CD5. Equivalent findings have not been reported previously in humans. We identified two discrete subsets of human CD8+ TN cells, defined by the absence or presence of the chemokine receptor CXCR3. The more abundant CXCR3+ TN cell subset displayed an effector-like transcriptional profile and expressed TCRs with physicochemical characteristics indicative of enhanced interactions with peptide-HLA class I Ags. Moreover, CXCR3+ TN cells frequently produced IL-2 and TNF in response to nonspecific activation directly ex vivo and differentiated readily into Ag-specific effector cells in vitro. Comparative analyses further revealed that human CXCR3+ TN cells were transcriptionally equivalent to murine CXCR3+ TN cells, which expressed high levels of CD5. These findings provide support for the notion that effector differentiation is shaped by heterogeneity in the preimmune repertoire of human CD8+ T cells.

Center for Cancer Research National Cancer Institute Bethesda MD 20892

Central European Institute of Technology 621 00 Brno Czech Republic

Department of Life Sciences University of Modena and Reggio Emilia 41125 Modena Italy

Department of Medical Biotechnologies and Translational Medicine University of Milan 20122 Milan Italy

Division of Genetic and Biomedical Research UoS Milan National Research Council 20089 Rozzano Milan Italy

Division of Immunology Transplantation and Infectious Diseases and Experimental Imaging Center IRCCS San Raffaele Scientific Institute and Vita Salute San Raffaele University 20132 Milan Italy

Division of Infection and Immunity Cardiff University School of Medicine Cardiff CF14 4XN United Kingdom

Genomic Unit Humanitas Clinical and Research Center 20089 Rozzano Milan Italy

Humanitas Flow Cytometry Core Humanitas Clinical and Research Center 20089 Rozzano Milan Italy

IFOM FIRC Institute of Molecular Oncology 20139 Milan Italy

Institute for Research in Biomedicine Faculty of Biomedical Sciences USI 6500 Bellinzona Switzerland

Institute of Microbiology ETH Zurich 8093 Zurich Switzerland

IRGB National Research Council 09042 Monserrato Italy

Laboratory of Molecular Immunology National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda MD 20892

Laboratory of Translational Immunology Humanitas Clinical and Research Center 20089 Rozzano Milan Italy

Laboratory of Translational Immunology Humanitas Clinical and Research Center 20089 Rozzano Milan Italy;

Pirogov Russian National Research Medical University 117997 Moscow Russia

Regensburg Center for Interventional Immunology University Regensburg and University Hospital Regensburg 93053 Regensburg Germany

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry 117997 Moscow Russia; and

Systems Immunity Research Institute Cardiff University School of Medicine Cardiff CF14 4XN United Kingdom

Unit of Clinical and Experimental Immunology Humanitas Clinical and Research Center 20089 Rozzano Milan Italy

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Functionally specialized human CD4+ T-cell subsets express physicochemically distinct TCRs