Figure 1.

Proliferating T cells following PD-1 ligation express diverse subset markers

(A) A description of the experimental system used for T cell stimulation.

(Bi) Primary human CD8 T cells were stimulated as indicated, with plate bound anti-CD3 and PD-L1 or PD-L2, and proliferation was measured by flow cytometry using celltrace far red proliferation dye dilution. Arrow indicates the cells that proliferated further.

(Bii) Quantification of the previous experiment, showing the percentage of proliferating cells. n = 4, one-way ANOVA, ∗∗∗p < 0.001, ns; not significant.

(C) The flow cytometry markers that were used to define the different cell subsets.

(D) CD8 T cells were stimulated as indicated and flow cytometry was used on day 4 to assess proliferation dye intensity and expression of subset specific markers. Arrows indicate the cell the proliferated further.

(Ei) Dot plot flow analysis of the previous experiment showing the percentages of each subset of T cells in the setting of the different stimulation conditions.

(Eii) Quantification of the percentage of the proliferating cells of same experiments, n = 4, two-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ns; not significant.

(F) A description of the experimental system used for T cell stimulation.

(G and H) Primary human CD3 T cells were stimulated as indicated and flow cytometry was used to asses proliferation and subset distribution.

(Ii) Dot plot flow analysis of CD4 T cells showing the percentages of each subset of T cells in the setting of the different stimulation conditions.

(Iii) Quantification of the percentage of the proliferating cells of same experiments, n = 3, two-way ANOVA, ns; not significant.

(Ji) Dot plot flow analysis of CD8 T cells showing the percentages of each subset of T cells in the setting of the different stimulation conditions.

(Jii) Quantification of the percentage of the proliferating cells of same experiments, n = 3, ns; not significant.