Figure 2.

Anti-CD3/CD28 stimulation activates all classes of lymphocytes. (A) Clustering of PBMCs at baseline and anti-CD3/CD28 conditions separates lymphocytes (NK, T, and B cells) primarily by cell type and secondarily by condition. Cells are color-coded based on activation condition (left) and cell types inferred via CITE-seq antibodies (right). (B) Annotation of cells at baseline was performed using protein expression levels of CD3, CD4, CD8, CD45RA, CD45RO, and additional annotation of activated T cells was done using expression of CD25 and CD69. (C) Cell type proportions across 10 donors at baseline (top) and anti CD3/CD28 condition (bottom). Note the decline in CD14⁺ monocytes (red bars) upon anti-CD3/CD28 condition. (D) Unsupervised clustering of all CD4⁺ T cells separates anti-CD3/CD28-stimulated cells from those at baseline and LPS activation. Dimension reduction plot with CD4⁺ T cells colored by their resultant cell type identity. (E) Average scaled expression of protein markers used to identify and annotate CD4⁺ T cell subsets. (F) Average scaled expression of marker genes associated with naive, MAIT, NK, TEMRA, and senescence functions.