Journal: Genes and immunity

Article Title: Despite IFN-lambda receptor expression, blood immune cells, but not keratinocytes or melanocytes, have an impaired response to type III interferons: implications for therapeutic applications of these cytokines.

doi: 10.1038/gene.2009.72

Figure Lengend Snippet: Figure 4 Peripheral blood mononuclear cells (PBMCs), keratino- cytes and HepG2 cells express interferon (IFN)-lR1 and IL-10R2 protein. Protein expressions of IFN-l receptor components and of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were analyzed in the cell lysates of PBMCs from three donors as well as of keratinocytes and HepG2 cells using the western blot technique.

Article Snippet: For the detection of IFN-l receptor components, we incubated blots with anti-IFNlR1 pAbs (50 ng ml 1; Sigma-Aldrich), anti-IL-10R2 pAbs (600 ng ml 1; R&D Systems) and anti-GAPDH mAb (200 ng ml 1; clone 6C5; Millipore), followed by peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H and L), rabbit anti-goat IgG (H and L) and goat antimouse IgG (H and L) (all from Dianova) incubation, respectively, and final ECL detection.

Techniques: Western Blot

Journal: Oncology reports

Article Title: Protective effect of neutralizing anti-IL-18α monoclonal antibody on a mouse model of acute graft-versus-host disease.

doi: 10.3892/or.2015.4176

Figure Lengend Snippet: Figure 1. Anti-IL-18Rα mAb alleviates aGVHD systemic symptoms in aGVHD mice. aGVHD was induced in irradiated BALB/c mice by BMC transplanta- tion. Ten micrograms of anti-IL-18Rα mAb was administered by intraperitoneal injection to mice in the BS+Ab group, while the BS group received injection of PBS. (A) Hunch posture, hair loss and skin lesions were evaluated on day 14 P.T. (B) Changes in body weight of the recipient mice at different time‑points in the different groups. (C) aGVHD clinical scores of the BS+Ab and BS groups on day 14 P.T. (D) Survival rates of aGVHD mice in the different groups. n=6 in each group. *P<0.05. GVHD, graft-versus-host disease; IL-18, interleukin-18; aGVHD, acute GVHD; BMC, bone marrow cell; PBS, phosphate-buffered solution; mAb, monoclonal antibody; P.T., post‑transplantation.

Article Snippet: Recipient mice in the BS+Ab group also received 10 μg/mouse intraperitoneal injection of neutralizing mAb against murine IL-18Rα (catalog no. MAB12161; R&D Systems, Minneapolis, MN, uSA) every 2 days.

Techniques: Irradiation, Injection

Journal: Oncology reports

Article Title: Protective effect of neutralizing anti-IL-18α monoclonal antibody on a mouse model of acute graft-versus-host disease.

doi: 10.3892/or.2015.4176

Figure Lengend Snippet: Figure 2. Effect of anti-IL-18Rα mAb administration on Th cell subsets, pro-inflammatory cytokines and histological scores in the aGVHD mice. (A-C) Peripheral blood levels of Th1 (A), Th2 (B) and Th17 (C) cell subsets in the BS+Ab and BS experimental groups were measured by flow cytometry at different time-points. Serum levels of IFN-γ (D), IL-4 (E), IL-17A (F) and IL-6 (H) at different time-points were detected by cytometric bead array, and IL-18 levels (G) were measured by ELISA. (I) Representative H&E staining of the liver and small intestine tissues of the mice in the BS+Ab and BS groups and the normal control group (untreated animals) on day 14 P.T. Magnification, x400. (J) Histological score was measured on day 14 P.T. n=6 in each group, *P<0.05. GVHD, graft-versus-host disease; aGVHD, acute GVHD; mAb, monoclonal antibody; Th, T helper; IL-18, interleukin-18; ELISA, enzyme-linked immunosorbent assay; H&E, hematoxylin and eosin; P.T., post‑transplantation.

Article Snippet: Recipient mice in the BS+Ab group also received 10 μg/mouse intraperitoneal injection of neutralizing mAb against murine IL-18Rα (catalog no. MAB12161; R&D Systems, Minneapolis, MN, uSA) every 2 days.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Control

Journal: Frontiers in oncology

Article Title: IL1 Pathway in HPV-Negative HNSCC Cells Is an Indicator of Radioresistance After Photon and Carbon Ion Irradiation Without Functional Involvement.

doi: 10.3389/fonc.2022.878675

Figure Lengend Snippet: FIGURE 4 | Evaluation of SASP gene expression after irradiation with 8 Gy photons or 4 Gy 12C-ions. (A) Schematic of experimental procedure. (B) Principal component analysis (PCA) on z-scaled data of the five cell lines for the 49 SASP factors. (C) Hierarchical clustering (fold change >1.5) for 11 genes to separate irradiated from unirradiated samples for photons and (D) 12C-ions. (E) Plot depicting fold change and q value for significantly differently expressed genes between unirradiated and irradiated samples for photons and 12C-ions. (F) IL1A and (G) IL1B values 72 h after irradiation were calibrated to the unirradiated control and correlated with radioresistance AUC or senescence AUC. (H) Correlation of IL1 pathway activity with radioresistance AUC or senescence AUC. Experiments were performed in three biol. replicates. Values are MV+/- SEM. T-test statistics were used for p value calculation. p < 0.05 are considered significant. R2: Pearson coefficient. For PCA and hierarchical clustering, log2-transformed, normalized (mean = 0, var = 1) data was used. Benjamini–Hochberg correction was applied, and q < 0.05 was considered significant. (I) Detection of IL1B protein secretion by ELISA up to 6 days after irradiation with 8 Gy photons in the cell lines UPCI:SCC131, UPCI:SCC040, and Cal33.

Article Snippet: Cytokine concentrations in cell culture supernatants were measured with the human IL1B DuoSet ELISA System (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Gene Expression, Irradiation, Control, Activity Assay, Transformation Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in oncology

Article Title: IL1 Pathway in HPV-Negative HNSCC Cells Is an Indicator of Radioresistance After Photon and Carbon Ion Irradiation Without Functional Involvement.

doi: 10.3389/fonc.2022.878675

Figure Lengend Snippet: FIGURE 5 | Cell survival after siRNA knockdown of ILA or/and IL1B. (A) Colony formation assay was initiated 1 day after transfection. Knockdown and control cells were irradiated with 2–8 Gy photons or 1–3 Gy 12C-ions. (B) Foci formation in response to 2-Gy photons or 1-Gy 12C-ions was examined by gH2AX/53BP1 immunofluorescence 24 h after irradiation. (C) Flow cytometry-based analysis of senescence after siRNA knockdown of IL1B in the cell line UPCI:SCC040. IL1B knockdown and control samples were irradiated with 6 Gy photon or 3 Gy 12C-ion, and senescence was determined from days 2 to 6 after irradiation. Experiments were performed in triplicates. MV+/- SEM are given. p values were calculated using t-test statistics. p = ns describes no significance was detected.

Article Snippet: Cytokine concentrations in cell culture supernatants were measured with the human IL1B DuoSet ELISA System (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Knockdown, Colony Assay, Transfection, Control, Irradiation, Flow Cytometry