Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: The expression of sIL-27Rα level in GVHD patients. ( A ) Serum sIL-27Rα levels were significantly increased on the day of neutrophil engraftment compared with pre-conditioning ( P < 0.0001). ( B ) The serum sIL-27Rα levels in patients with grade II–IV aGVHD were significantly lower than those of 0-I aGVHD patients on the day of neutrophil engraftment ( P < 0.001). ( C ) On the day of neutrophil engraftment, serum sIL-27Rα levels in patients with skin aGVHD ( P < 0.01) and liver aGVHD ( P < 0.05) were significantly lower than those of 0-I aGVHD patients. ( D ) There were no similar significant difference regarding cGVHD. Data shown are mean ± SD.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: The diagnostic value of sIL-27Rα in aGVHD and the association of sIL-27Rα with aGVHD severity, relapse and survival. ( A ) The area under the ROC curve (AUC) was 0.735 (95% CI 0.618–0.853, P = 0.001) on the day of neutrophil engraftment; when using 59.4 ng/ml as cut-off value from the ROC curve, the sensitivity and specificity were 56% and 81%, respectively. ( B ) The cumulative incidence of grade II–IV aGVHD was significantly lower in patients with high sIL-27Rα levels by Gray’s test ( P = 0.004). ( C ) Patients with high sIL-27Rα levels showed favourable overall survival compared with patients with low sIL-27Rα levels with Kaplan-Meier survival analysis by log rank test ( P < 0.001). ( D , E ) Patients with high sIL-27Rα levels had lower relapse rate (CIR) and non-relapse mortality (NRM) than did patients with low sIL-27Rα levels by Gray’s test (P = 0.008, respectively).

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Diagnostic Assay

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: The association of sIL-27Rα levels at neutrophil engraftment with clinical factors.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Biomarker Discovery

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: Univariate and multivariate analyses of factors affecting the incidence of grade II–IV acute graft- versus-host disease after allogeneic hematopoietic stem cell transplantation at neutrophil engraftment.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Transplantation Assay

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: sIL-27Rα as an independent prognostic aGVHD biomarker validated in an independent cohort of 80 patients. ( A ) The area under the ROC curve (AUC) was 0.790 (95% CI 0.688–0.892, P < 0.001) on the day of neutrophil engraftment, using 59.4 ng/ml as the cut-off value. ( B ) The cumulative incidence of grade II–IV aGVHD was significantly lower in patients with high sIL-27Rα levels by Gray’s test ( P < 0.001). ( C ) Patients with high sIL-27Rα levels showed favourable overall survival compared with patients with low sIL-27Rα levels on Kaplan-Meier survival analysis by log rank test ( P = 0.012). ( D , E ) Patients with high sIL-27Rα levels had similar relapse rates (CIR) and lower non-relapse mortality (NRM) than did patients with low sIL-27Rα levels by Gray’s test ( P = 0.0931, P = 0.005, respectively).

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Biomarker Discovery

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: Association of sIL-27Rα with serum cytokine levels in aGVHD. The associations between sIL-27Rα levels with serum IL-27, IL-10, HGF, TNFR1, elafin, REG3α and ST2 levels were analysed by using Spearman’s rank correlation coefficient.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques:

Journal: Mediators of Inflammation

Article Title: IL-27 Activates Human Trophoblasts to Express IP-10 and IL-6: Implications in the Immunopathophysiology of Preeclampsia

doi: 10.1155/2014/926875

Figure Lengend Snippet: Analysis of IL-27 and IL-27 receptor expression in placental tissues. (a) Representative Western blot analysis of IL-27 receptor subunits WSX-1 in placental tissues. The first three are representative of samples of normal pregnancy (N) group and the last is from the PE group. (b)–(i) Immunohistochemical staining of IL-27 and WSX-1 in the placental tissues. (b) and (d) show the immunostaining of IL-27 in normal control (N) group; (c) and (e) show the immunostaining of IL-27 in preeclampsia (PE) group. (f) and (h) show the immunostaining of the WSX-1 in normal control (N) group; (g) and (i) show the immunostaining of WSX-1 in preeclampsia (PE) group. β -Actin was used as protein control to ensure an equal amount of loaded protein. Original magnification: 100x for c, f, g, and h; 400x for d, e, h, and i. All experiments were performed in three independent replicates.

Article Snippet: Sections were incubated with anti-IL-27 mAb (Abcam115671, HK) and anti-WSX-1 mAb (RD AF1479), diluted in 5% (wt/vol) nonfat milk for 16 h at 4°C.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining, Immunostaining

Journal: Mediators of Inflammation

Article Title: IL-27 Activates Human Trophoblasts to Express IP-10 and IL-6: Implications in the Immunopathophysiology of Preeclampsia

doi: 10.1155/2014/926875

Figure Lengend Snippet: IL-27 and IL-27 receptor was expressed in HTR-8/SVneo cells and it upregulated CXCL10 and IL-6. (a) RNA and protein expression for WSX-1. (b) RNA and protein expression of gp130. From the left to the right lane, respectively, are Hct116 (human colon cancer cells), Skov3 (ovarian carcinoma cells), and HTR-8/SVneo. Hct116 and Skov3 were used as control. GAPDH was used as protein control to ensure an equal amount of loaded protein. (c) Effects of IL-27 (50 ng/mL) on the mRNA expression of inflammatory mediators. Kinetic gene expressed of CXCL10 (d) and IL-6 (e) in HTR-8/SVneo cells under stimulated with IL-27. * P < 0.05, ** P < 0.01, and *** P < 0.001 when compared between groups denoted by horizontal lines ( n = 3). (f) CXCL10 protein levels after the stimulation of IL-27 (0–50 ng/mL) in cell supernatant of HTR-8/SVneo (0–30 h). (g) IL-6 protein expression after the stimulation of IL-27 (0–50 ng/mL) in cell supernatant of HTR-8/SVneo (0–48 h). * P < 0.05, ** P < 0.01, and *** P < 0.001 when comparing the level at the starting time point. All experiments were performed in three independent replicates.

Article Snippet: Sections were incubated with anti-IL-27 mAb (Abcam115671, HK) and anti-WSX-1 mAb (RD AF1479), diluted in 5% (wt/vol) nonfat milk for 16 h at 4°C.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Interleukin-27-polarized HIV-resistant M2 macrophages are a novel subtype of macrophages that express distinct antiviral gene profiles in individual cells: implication for the antiviral effect via different mechanisms in the individual cell-dependent manner

doi: 10.3389/fimmu.2025.1550699

Figure Lengend Snippet: Quantification of biomarkers.

Article Snippet: The following antibodies and their respective fluorochrome were used: CD38 (Fluorochrome PE-Cy7; Cat. # 560677, BD Biosciences, Franklin Lakes, NJ, USA), isotype control (Fluorochrome PE-Cy7; Cat. # 557872, BD Biosciences), CD130/GP130 (Fluorochrome AF647, Cat. # 564151, BD Biosciences), isotype control (Fluorochrome AF647, Cat. # 557714, BD Biosciences), WSX1 (Fluorochrome PE; Cat. # FAB14791PO2, R&D Systems), and isotype control (Fluorochrome PE; Cat. # IC0041P, R&D Systems).

Techniques:

Journal: Frontiers in Immunology

Article Title: Interleukin-27-polarized HIV-resistant M2 macrophages are a novel subtype of macrophages that express distinct antiviral gene profiles in individual cells: implication for the antiviral effect via different mechanisms in the individual cell-dependent manner

doi: 10.3389/fimmu.2025.1550699

Figure Lengend Snippet: Detection of WSX and gp130 on M2 macrophages using FACS. The surface expression of WSX1 and gp130 on M2 macrophages was assessed by flow cytometry as described in the Materials and Methods. (A) The left and right panels show WSX1 and gp130 (CD130) staining, respectively. The staining pattern of isotype control antibodies is shown in gray line, and black indicates the protein of interest. The x-axis and y-axis show fluorescence intensity and cell counts, respectively. The percentages in the panels indicate the percentage of cells expressing WSX1 or gp130 in the samples. Data are representative of three independent experiments with similar outcomes. (B) Flow cytometric analysis of M2 macrophages showing the expression of WSX1 and gp130 (CD130) from one donor M2 macrophages. (C) FACS analysis for WSX1 and gp130 expression on M2 cells was performed using three independent donor cells. Results indicated means ± SE (n = 3). FACS, fluorescence-activated cell sorting.

Article Snippet: The following antibodies and their respective fluorochrome were used: CD38 (Fluorochrome PE-Cy7; Cat. # 560677, BD Biosciences, Franklin Lakes, NJ, USA), isotype control (Fluorochrome PE-Cy7; Cat. # 557872, BD Biosciences), CD130/GP130 (Fluorochrome AF647, Cat. # 564151, BD Biosciences), isotype control (Fluorochrome AF647, Cat. # 557714, BD Biosciences), WSX1 (Fluorochrome PE; Cat. # FAB14791PO2, R&D Systems), and isotype control (Fluorochrome PE; Cat. # IC0041P, R&D Systems).

Techniques: Expressing, Flow Cytometry, Staining, Control, Fluorescence, FACS