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. 2021 Jan 8;11:577655. doi: 10.3389/fimmu.2020.577655

Figure 5.

Figure 5

T cell expressed CD80 inhibits CTLA-4 availability and promotes Treg growth. (A) CD4+CD25- T cells were stimulated with CD3/CD28 beads in presence of anti-CTLA-4 and the expression of CD80 and CD86 analysed five days post stimulation by flow cytometry. Representative FACS plots and pooled data for percentage of CD80 and CD86 expressing cells in gated divided cells are shown. P-values were calculated using paired two-tailed T-test. (B) CD4+CD25- T cells were stimulated for five days with CD3/CD28 beads. On day four, anti-CD80 was added to culture overnight, and CTLA-4 expression was measured by flow cytometry the following day. Representative histogram and collated data of CTLA-4 expression in the presence or absence of anti-CD80. P-values were calculated using paired two-tailed T-test. (C) Jurkat-CTLA-4 cells were transduced with CD80GFP and the level of cycling CTLA-4 was examined by flow cytometry. Cells were cultured overnight in the presence or absence of anti-CD80 where indicated. Representative histogram and collated data of CTLA-4 expression. P-values were calculated using paired two-tailed T-test. (D) Expanded Tregs were restimulated with CHO-CD86 or anti-CD3/anti-CD28 beads in the presence or absence of anti-CD80, anti-CTLA-4 or both. Boxplots show the total number of Tregs following treatment. P-values were calculated using one-way ANOVA. n.s., not significant.