Figure 2. A subpopulation of Tregs in the peripheral blood of sJIA patients share a mass cytometric phenotype with sJIA synovial fluid Tregs.

(A–H) Mass cytometry data of Tregs (CD4⁺CD25⁺CD127^lo) were concatenated by study subject group, chronic sJIA SF (n = 2), acute sJIA PB (n = 6), chronic sJIA PB (n = 4), and controls (n = 6), and evaluated by viSNE and SPADE. (A–D) viSNE plots of mass cytometry of Tregs are depicted. The black circles on the viSNE plots in B and C highlight a population of Tregs that are cytometrically similar to SF Tregs depicted in A based on the location of the cells on the viSNE plot. The cell markers expressed in the population of Tregs identified on the viSNE plots by the black circle are shown in the accompanying bar graphs. (D) For healthy controls who lacked this population of Tregs, the expression of cell markers in all Tregs on the viSNE plot is depicted. (E–H) SPADE visualization of the same data. The size of the nodes reflects cell number, and the color of the node reflects the percent of total events. The bubbled nodes (black circle) on the SPADE trees captures the dominate population of Tregs in sJIA SF. The cell markers expressed on Tregs in the SPADE bubble are shown in the accompanying bar graphs. (H) For healthy controls who had a small number of cells in the SPADE bubble, the expression of cell markers on Tregs outside of this bubble are depicted. viSNE and SPADE analyses were performed with Cytobank. sJIA, systemic juvenile idiopathic arthritis; PB, peripheral blood; SF, synovial fluid; viSNE, visualization using t-Distributed Stochastic Neighbor Embedding; tSNE, t-Distributed Stochastic Neighbor Embedding; SPADE, spanning-tree progression analysis of density-normalized events.