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Figure 2 | Journal of Translational Medicine

Figure 2

From: Complement component C5a Promotes Expression of IL-22 and IL-17 from Human T cells and its Implication in Age-related Macular Degeneration

Figure 2

IL-1β and IL-6 secreting monocytes are important for C5a induced IL-22 and IL-17 expression form T cells. (A) CD3+CD4+ T (T) cells and CD3-CD14+ monocytes (M) were sorted and cultured with or without C5a for 3 days. Cell supernatants were assessed for IL-22 and IL-17 expression. Three separate experiments were performed and the figure shows representative data. (B) CD3+CD4+CD45RA+ (naïve T cells, nT) and CD3+CD4+CD45RA- (memory T cells, mT) T cells and CD3-CD14+ monocytes (M) were sorted and cultured with or without C5a for 3 days. IL-22 and IL-17 levels were measured from supernatants. Three separate experiments were performed and the figure shows representative data. (C) C5a activates B7 expression on monocytes. PBMCs were cultured with or without C5a for 1 day. CD3-CD14+ monocytes were gated for indicated cell markers' expression. Similar results were seen in another independent assay. (D) IL-22 and IL-17 in 3-day culture supernatants of PBMCs with the presence or absence of C5a, C5aR antagonist and anti-B7.1 and anti-B7.2 antibodies. (E) C5a stimulates monocytes to secrete IL-1β and IL-6. PBMCs were cultured with or without C5a and C5aR antagonist for 3 days. Cell supernatants were assayed for IL-1β, IL-6 and TNFα expression. (F) Monocytes and T cells were sorted and cultured with or without C5a for 3 day. Cell supernatants were assayed for IL-1β and IL-6 expression. Three separate experiments were performed and the figure shows representative data. (G) IL-22 and IL-17 in 3-day culture supernatants of PBMCs with the presence or absence of C5a with isotype control antibody, C5aR antagonist and anti-IL-1β and anti-IL-6 neutralization antibodies. Three separate experiments were performed.

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