Fig. 2.

Stimulation of CD4⁺ T lymphocytes with TGF-β1 and TGF-β3 upregulates CD70, and CD70 engagement promotes proinflammatory cytokine expression. a Proportion of CD70⁺CD4⁺ T lymphocytes after in vitro stimulation with α-CD3/α-CD28 alone or together with various cytokines (α-CD3/α-CD28 used as baseline) in healthy controls (n = 7). b Percentage of CD70⁺ lymphocytes within IFN-γ⁻IL-17⁺ producing CD4⁺ T lymphocytes after in vitro stimulation with α-CD3/α-CD28 alone or together with various cytokines in healthy controls (n = 7). c Quantitative PCR analysis of granzyme B (Grz B), IL-23 receptor (IL23-R), FOXP3, ROR-γt, GATA3, and T-bet mRNA in CD70⁺CD4⁺ T lymphocytes and CD70^negCD4⁺ T lymphocytes after in vitro stimulation with TGF-β1. The results are shown as ratios (CD70⁺/CD70^neg) of mRNA transcripts relative to 18S ribosomal RNA, in healthy controls (n = 5). d Relative expression of IL-17, IFN-γ, and GM-CSF within CD4⁺ T lymphocytes (stimulated with TGF-β1) incubated with or without soluble CD27 (sCD27, 8 ng/ml) for 3 days in healthy controls (n = 6). The results are shown as the ratio (addition of soluble CD27/no addition of soluble CD27). e Representative dot plot of CD70 expression on IL-17, IFN-γ, and GM-CSF-expressing CD4⁺ T lymphocytes after in vitro stimulation with α-CD3/α-CD28 alone or together with TGF-β1 or TGF-β3 in a healthy control (representative of n = 6). f, g Percentage of IFN-γ⁺IL-17⁺ and IL-17⁺GM-CSF⁺ in CD70⁺ and CD70^neg CD4⁺ T lymphocytes after in vitro stimulation with α-CD3/α-CD28 alone or together with TGF-β1 or TGF-β3 in healthy controls (n = 6). A one-way ANOVA was used to compare various stimulation conditions and a paired t-test to compare CD70⁺ with CD70^neg and TGF-β1 with TGF-β3. Data can be expressed relative to control (α-CD3/α-CD28), with or without the addition of soluble CD27, and relative to CD70⁺. Statistical analysis was performed on raw data. Data are represented as the mean ± SEM *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001