Fig. 3.
Macrophages cultured with SSc patient fibroblast-derived exosomes upregulate pro-inflammatory and pro-fibrotic cytokines
Human monocytes were cultured in complete media with 20 ng/ml M-CSF for 5 days. On the 5th day, cells were washed and complete media with exosome-depleted FBS were added to macrophages prior to treatment with healthy or SSc-fibroblast-derived exosomes for 48 h. At the conclusion of activation, macrophage supernatants were collected and analysed by multiplex for secreted protein production. Release of IL-1α (A), IL-6 (B), IL-10 (C), IL-12 (p40) (D), IL-22 (E), TGF-α (F), TNF (G), TNF-β (lymphotoxin) (H), VEGF-A (I) and CCL2 (J) were increased by macrophages stimulated with SSc patient fibroblast-derived exosomes compared with healthy control fibroblast-derived exosomes. *P < 0.05, **P < 0.01; data shown are representative of results obtained from analysis of supernatant collected from macrophages stimulated with fibroblast-derived exosomes from 4 healthy donors or 5 SSc patients. Error bars represent s.d.