TCR signal strength shapes functional imprinting during CD4 T cell development

Abstract

Abstract Naïve CD4 and CD8ab T cells are selected in the thymus in response to low-avidity TCR interactions. Such weak TCR signals install a quiescent program that guides naïve T cells to the lymph nodes where they differentiate upon encounter of cognate antigen. Thymocytes expressing self-specific TCRs can be agonist-selected in response to a strong TCR signal and differentiate in the thymus to functionally committed double negative T cells, NKT cells or regulatory T cells. Although the role for strength of TCR activation is clear in instructing these different outcomes, the impact of self-recognition in the thymus on “naïve” T cells is unclear. Previous work demonstrated that negative selection of self-specific T cells is not all encompassing and that affinity for self could even enhance immune protection against pathogens. We have used Stim1fl/fl Stim2fl/fl conditional deletion mice that fail to sustain calcium entry into the cytosol upon TCR activation to assess the role of TCR strength during selection. We confirmed that agonist selection is drastically affected in these mice whereas numbers of conventional T cells are not. Interestingly, viSNE representation of multi-parameter flow cytometry revealed single cell heterogeneity among the CD4SP thymocyte subset that is controlled by Stim1/Stim2. This indicates that differential TCR activation levels give rise to phenotypically different CD4 T cells. Purification of a strongly signaled (Stim1/2-dependent) subset of CD4 thymocytes showed functional differences in terms of their ability to proliferate in response to TCR activation. Our work points to a role for self-recognition during T cell selection that shapes the response of individual CD4 T cells to TCR activation in the periphery.