Figure 2. Reduced IL-6 production contributes to impaired Th17 polarization in tolerant recipients.

(A) Representative dot plots and (B) total percentage of CD14, CD14⁺⁺CD16^-, CD14⁺⁺CD16⁺ and CD14⁺CD16⁺⁺ monocytes in peripheral blood samples from patients and healthy volunteers. Statistical significance was determined using Kruskal-Wallis ANOVAs with Dunn’s multiple comparison tests. (C) HLA-DR, CD86 and CD80 MFI were measured in monocyte subsets from patients and healthy volunteers. Statistical significance was determined using Repeated Measurement (RM) 2-Way ANOVAs with Sidak’s multiple comparison tests. (D) IL-6 and TNF-α production by 1.25x10⁵ monocytes isolated from patients and healthy volunteers was measured after LPS activation (50ng/ml). Statistical significance was determined using RM 2-Way ANOVAs with Sidak’s multiple comparison tests. (E) CD163 expression was measured in monocyte subsets from patients and healthy volunteers. Statistical significance was determined using RM 2-Way ANOVAs with Sidak’s multiple comparison tests. (F) IL-6, TNF-α and IL-17 production was measured in cocultures consisting of 5x10⁴ CD4⁺T cells and 2.5x10⁴ monocytes in the presence or absence of soluble 10ng/mL anti-CD3, using cells isolated from patients and healthy volunteers. Statistical significance was determined using RM 2-Way ANOVAs with Sidak’s multiple comparison tests. Statistical significance is denoted as follows: ****=P<0.0001, ***=P<0.001, **=P<0.01 and *=P<0.05.