Figure 5
GrzB production by memory CD4 T cell subsets and coexpression with cytokines. (A-B) GrzB expression by Tregs. Memory CD4 T cells were purified from peripheral blood, and 5×105 cells (in 1ml medium) were untreated (UT) or activated (CD3/CD28 costimulation) for 24 hrs. Cells were then stained for CD25, Foxp3, and GrzB. (A) CD25 expression levels (Negative, Dim, or Bright) by memory CD4 T cells after 24 hrs culture (mean ± sem, n = 3). (B) Foxp3/GrzB expression by CD25-Negative, CD25-Dim, or CD25-Bright memory CD4 T cells after 24 hrs culture. Graph shows the mean (n = 3) distribution of Foxp3/GrzB subsets gated on CD25-Negative, -Dim, or -Bright cells. (C) GrzB coexpression with IFNγ, IL4, or IL17A by memory CD4 T cells. Memory CD4 T cells (5×105 cells in 1ml medium) were untreated for activated (CD3/CD28 costimulation or PMA/IO in the presence of brefeldin) for 24 hrs, then stained for GrzB and either IFNγ, IL4, or IL17A. Graphs show the mean (n = 3) distribution of cytokine/GrzB subsets. (D) GrzB/IFNγ coexpression by memory CD8 T cells. Purified memory CD8 T cells (5×105 cells in 1 ml medium) were untreated or activated (CD3/CD28 costimulation or PMA/IO in the presence of brefeldin) for 24 hrs. Graph shows the mean (n = 3) distribution of IFNγ/GrzB subsets.