Fig. 3

Influence of JAK, STAT5, and PI3K inhibitor to IL-7-mediated CD127 expression in CD8⁺ T cells in patients with primary cutaneous melanoma. 10⁴ of purified peripheral CD8⁺ T cells from fourteen patient with primary cutaneous melanoma were stimulated with recombinant human IL-7 (10 ng/mL) for 48 h in the presence of anti-CD3/CD28, along with either JAK inhibitor (10 µmol/L), STAT5 inhibitor (250 µmol/L), or PI3K inhibitor (25 µmol/L). Control cells were cultured with anti-CD3/CD28 only. Cells and supernatants were harvested. Total CD127 mRNA and sCD127 mRNA level in CD8⁺ T cells were measured by real-time PCR. a IL-7 stimulation promoted total CD127 mRNA expression in CD8⁺ T cells. Either JAK inhibitor or STAT5 inhibitor did not affect IL-7-mediated elevation of total CD127 mRNA expression, while PI3K inhibitor abrogated the effect of IL-7 on total CD127 mRNA induction in CD8⁺ T cells. b IL-7 stimulation also enhanced sCD127 mRNA level in CD8⁺ T cells. Either JAK inhibitor or STAT5 inhibitor did not influence IL-7-induced elevation of sCD127 mRNA expression, while PI3K inhibitor abrogated the effect of IL-7 on sCD127 mRNA induction in CD8⁺ T cells. c sCD127 level in the supernatants was measured by ELISA. IL-7 stimulation promoted sCD127 level in the cultured supernatant of CD8⁺ T cells. Either JAK inhibitor or STAT5 inhibitor did not influence IL-7-induced sCD127 secretion, while PI3K inhibitor abrogated the effect of IL-7 on sCD127 production by CD8⁺ T cells. Individual level of each subject is shown. Statistical analysis was performed using one-way ANOVA and LSD-t test