Table 1.
Demographic and clinical characteristics of patients.
Fig 1.
Anti-TNF-α plasma level and anti-drug antibodies.
(A) Adalimumab concentration in plasma from patients with psoriasis who respond (R) or do not respond (NR) to therapy. *p<0.05. (B) Etanercept concentration in plasma from patients with psoriasis who respond (responders, R) or do not respond (non responders, NR) to therapy. *p<0.05. (C) Infliximab concentration in plasma from R or NR to therapy. (D) Anti-infliximab antibodies concentration in plasma from R (n = 3) or NR (n = 3) to therapy. (E) Number of patients with anti-infliximab antibodies stratified according to infliximab plasma levels.
Fig 2.
Patients treated with etanercept have higher plasma levels of TNF-α.
(A) TNF-α concentrations in plasma from healthy controls (HC), and from psoriatic patients who were R or NR to adalimumab, etanercept or infliximab. The number of patients (n) was 29 for R to adalimumab, 16 for NR to adalimumab, 24 for R to etanercept, 12 for NR to etanercept, 30 for R to infliximab, 11 for NR to infliximab, 20 for HC. *p<0.05; **p<0.01; ***p<0.001. (B) sTNFRI concentrations in plasma from HC, and from psoriatic patients who were R or NR to adalimumab, etanercept or infliximab. The number of patients for each group is the same as in panel A. (C) sTNFRII concentrations in plasma from HC, and from psoriatic patients who were R or NR to adalimumab, etanercept or infliximab. The number of patients for each group is the same as in panel A. (D) Linear regression analysis for the correlation between plasma levels of TNF-α and plasma levels of sTNFRI. (E) Linear regression analysis for the correlation between plasma levels of TNF-α and plasma levels of sTNFRII, (F) Linear regression analysis for the correlation between plasma levels of sTNFRI and plasma levels of sTNFRII. (G) Linear regression analysis for the correlation between age and plasma levels of TNF-α. (H) Linear regression analysis for the correlation between age and plasma levels of sTNFRI. (I) Linear regression analysis for the correlation between age and plasma levels of sTNFRII.
Fig 3.
Principal component analysis bi-plot (loadings and scores).
The spatial distribution of the immunological parameters and of R and NR to different treatments is based on the first three principal components (PC). Each dot represents a single patient. Confidential ellipses are also shown. TNFA, plasma levels of TNF-α; TNFRI, plasma levels of sTNFRI; TNFRII, plasma levels of sTNFRII; ADA, plasma levels of adalimumab; ETA, plasma levels of etanercept; INFLIXI, plasma levels of infliximab.
Fig 4.
PBMCs from patients responding to adalimumab and etanercept produce higher levels of TNF-α and sTNFRII than PBMCs from patients responding to infliximab.
(A) TNF-α levels were quantified in the supernatant of peripheral blood mononuclear cells (PBMCs) in the absence or in the presence of lipopolysaccharide (LPS) for 24 hours. PBMCs were obtained from patients treated with adalimumab, etanercept or infliximab. Ada, adalimumab; Eta, etanercept; Inf, infliximab; NR, non-responders; R, responders. *p<0.05; ***p<0.001. (B) sTNFRI levels were quantified in the supernatant of PBMCs in the absence or in the presence of LPS for 24 hours. PBMCs were obtained from patients treated with Ada, Eta or Inf. *p<0.05. (C) sTNFRII levels were quantified in the supernatant of PBMCs in the absence or in the presence of LPS for 24 hours. PBMCs were obtained from patients treated with Ada, Eta or Inf. *p<0.05; **p<0.01; ***p<0.001.
Fig 5.
Gating strategy used to identify monocyte subsets among PBMCs.
Monocytes were first selected on the basis of their physical parameters, i.e. forward scatter (FSC) and side scatter (SSC). Doublets were eliminated from the analysis (upper right panel). Then, living monocyes were selected according to the negativity to Live Dead probe (middle left panel). Cells negative for the DUMP channel (anti-CD3, anti-CD19 and anti-CD56) were then selected (middle right panel). Among these, monocytes were selected according to positivity to CD14 and HLA-DR (lower left panel). In this population, three monocyte subsets were identified on the basis of the expression of CD14 and CD16 (lower right panel).
Fig 6.
Adalimumab affects the distribution of monocyte subsets.
(A) Percentage of CD14+ monocytes (i.e., all monocytes) in patients treated with adalimumab (Ada), etanercept (Eta) or infliximab (Inf). *p<0.05. (B) Quantification of HLA-DR expression in CD14+ monocytes. (C) Percentage of CD14++,CD16- (classical) monocytes in patients treated with Ada, Eta or Inf. *p<0.05. (D) Quantification of HLA-DR expression in CD14++,CD16- monocytes. (E) Percentage of CD14+,CD16++ (non-classical) monocytes in patients treated with Ada, Eta or Inf. (F) Quantification of HLA-DR expression in CD14+,CD16++ monocytes. (G) Percentage of CD14++,CD16+ (intermediate) monocytes in patients treated with Ada, Eta or Inf. (H) Quantification of HLA-DR expression in CD14++,CD16+ monocytes. For all panels: R, responders; NR, non-responders; MFI, median fluorescence intensity.