Fig. 4: T-cell activation is reduced by HARS-related proteins.

T cells isolated from PBMCs obtained from healthy volunteers were stimulated with plate-bound anti-CD3/anti-CD28 for 24 h in the presence of vehicle or recombinant human HARS.a IL-2 release by individual donors (n = 8) was measured, [HARS] = 0.3 nM. b IL-2 reduction by treatment with the vehicle control (n = 3) was measured after treatment with HARS at different concentrations. c Effect of HARS (0.3 nM) on the release of various cytokines and granzyme B (n = 2 donors in two separate experiments) were measured. d Flow cytometry analysis of CD40L expression on CD4 + T cells was performed, and the representative histogram and % positive results from individual donors (n = 9) are shown, [HARS] = 1 nM. e Flow cytometry analysis of CD69 expression on CD4 + (left) and CD8 + (right) T cells was performed, and representative scatter plots and MFI results from individual donors (n = 9) are shown, [HARS] = 1 nM. Error bars indicate SEM, paired t-test or ANOVA with Dunnett’s post hoc test where appropriate. *p < 0.05; **p < 0.01; ***p < 0.001, ****p < 0.0001.