Figure 5.

Acan1 and Nak1 modulate cytokine production in human immune cells.A, apoptosis population percentage in human peripheral blood mononuclear cells when incubated with synthetic hookworm peptides (10 μg/ml). Peptides were incubated for 4 h at 37 °C/10% CO₂. Heat-treated positive control was placed in a heating block for 20 min at 56 °C. Apoptosis was measured using Annexin V-FITC and propidium iodide staining and analyzed by flow cytometry. B, culture supernatant concentrations of TNF and IL-2 assessed by cytometric bead array from human peripheral blood mononuclear cells stimulated with phorbol 12-myristate 12-acetate (50 ng/ml) + ionomycin (1 μg/ml) (PI) for 24 h. Data show mean ± SEM of a representative experiment out of three, with n = 3. Unpaired Student t test performed comparing PI versus Acan1 or Nak1 + PI. C, CD4⁺ and CD8⁺ T cell proliferation percentage assessed by CTV assay from MACS-purified CD3⁺ human T cells stimulated with anti-CD3/CD28 beads and incubated with hookworm peptide (100 μg/ml) for 4 days at 37 °C/10% CO₂ before being analyzed by flow cytometry. Ciclosporin (10 μg/ml) was used as a positive control. Data show mean ± SEM of a representative experiment, n = 3. Unpaired Student t test was performed comparing anti-CD3/CD28 beads with Acan1 or Acan1 (linear); ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗p < 0.0001.