A CD4⁺ T cell reference map delineates subtype-specific adaptation during acute and chronic viral infections

CD4⁺ T cells are critical orchestrators of immune responses against a large variety of pathogens, including viruses. While multiple CD4⁺ T cell subtypes and their key transcriptional regulators have been identified, there is a lack of consistent definition for CD4⁺ T cell transcriptional states. In addition, the progressive changes affecting CD4⁺ T cell subtypes during and after immune responses remain poorly defined. Using single-cell transcriptomics, we characterized the diversity of CD4⁺ T cells responding to self-resolving and chronic viral infections in mice. We built a comprehensive map of virus-specific CD4⁺ T cells and their evolution over time, and identified six major cell states consistently observed in acute and chronic infections. During the course of acute infections, T cell composition progressively changed from effector to memory states, with subtype-specific gene modules and kinetics. Conversely, in persistent infections T cells acquired distinct, chronicity-associated programs. By single-cell T cell receptor (TCR) analysis, we characterized the clonal structure of virus-specific CD4⁺ T cells across individuals. Virus-specific CD4⁺ T cell responses were essentially private across individuals and most T cells differentiated into both Tfh and Th1 subtypes irrespective of their TCR. Finally, we showed that our CD4⁺ T cell map can be used as a reference to accurately interpret cell states in external single-cell datasets across tissues and disease models. Overall, this study describes a previously unappreciated level of adaptation of the transcriptional states of CD4⁺ T cells responding to viruses and provides a new computational resource for CD4⁺ T cell analysis.