Determination of Tr1 cell populations correlating with distinct activation states in acute IAV infection

Abstract

Type I regulatory (Tr1) cells are defined as FOXP3^-IL-10-secreting clusters of differentiation (CD4⁺) T cells that contribute to immune suppression and typically express the markers LAG-3 and CD49b and other co-inhibitory receptors. These cells have not been studied in detail in the context of the resolution of acute infection in the lung. Here, we identify FOXP3^- interleukin (IL)-10⁺ CD4⁺ T cells transiently accumulating in the lung parenchyma during resolution of the response to sublethal influenza A virus (IAV) infection in mice. These cells were dependent on IL-27Rα, which was required for timely recovery from IAV-induced weight loss. LAG-3 and CD49b were not generally co-expressed by FOXP3^- IL-10⁺ CD4⁺ T cells in this model and four populations of these cells based on LAG-3 and CD49b co-expression were apparent [LAG-3^-CD49b^- (double negative), LAG-3⁺CD49b⁺ (double positive), LAG-3⁺CD49b^- (LAG-3⁺), LAG-3^-CD49b⁺ (CD49b⁺)]. However, each population exhibited suppressive potential consistent with the definition of Tr1 cells. Notably, differences between these populations of Tr1 cells were apparent including differential dependence on IL-10 to mediate suppression and expression of markers indicative of different activation states and terminal differentiation. Sort-transfer experiments indicated that LAG-3⁺ Tr1 cells exhibited the capacity to convert to double negative and double positive Tr1 cells, indicative of plasticity between these populations. Together, these data determine the features and suppressive potential of Tr1 cells in the resolution of IAV infection and identify four populations delineated by LAG-3 and CD49b, which likely correspond to different Tr1 cell activation states.