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: Stem Cell Reports

Article Title: IL-18R-mediated HSC quiescence and MLKL-dependent cell death limit hematopoiesis during infection-induced shock

doi: 10.1016/j.stemcr.2021.10.011

Figure Lengend Snippet: IL-18R blockade preserves ST-HSCs and mitigates quiescence WT (C57BL/6) mice were inoculated with 10 5 copies of IOE and BM was analyzed at 7 dpi. (A) Model depicting treatment of WT mice with either IL-18Rα blocking antibody or an isotype control. (B–E) Absolute cellularity of (B) BM, (C) HSPCs, (D) LT-HSCs, and (E) ST-HSCs at 7 dpi, n = 3–8 mice/group. (F and G) The frequency of quiescent (F) LT-HSCs and (G) ST-HSCs, n = 3–8 mice/group. (H) Bacterial burden in the spleen of IOE-infected mice treated with either isotype or anti-IL-18Rα, quantified by copies of the dsb gene. (I and J) Cell death breakdown of (I) LT-HSC and (J) ST-HSCs stained with 7AAD and Annexin V in mock- or IOE-infected WT mice treated with mock, isotype control, or anti-IL18Rα, n = 7–9 mice/group. ∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗ p < 0.0001.

Article Snippet: Mice were administered 75 μg/mouse of anti-IL-18Rα antibodies (clone AF856; R&D Systems) or IgG2 isotype control antibody via i.p. injection on days 4 and 6 post-infection.

Techniques: Blocking Assay, Control, Infection, Staining

Journal: Stem Cell Reports

Article Title: IL-18R-mediated HSC quiescence and MLKL-dependent cell death limit hematopoiesis during infection-induced shock

doi: 10.1016/j.stemcr.2021.10.011

Figure Lengend Snippet: IL-18R directly regulates quiescence and indirectly induces cell death WT hosts were lethally irradiated and transplanted with an equal ratio of WT (GFP) and Il18r −/− BM cells (BMCs). (A) Model for the generation of WT: Il18r −/− mixed chimeras. Mice were inoculated with 10 5 copies of IOE and BM was analyzed at 7 dpi. (B–D) The absolute cellularity of (B) HSPCs, (C) LT- HSCs, and (D) ST-HSCs of either WT or Il18r −/− donor origin in the BM of WT recipients, n = 5–7 mice/group. (E and F) The frequency of quiescent (E) LT-HSCs and (F) ST-HSCs in a mixed BM chimera setting. (G–I) Cell death breakdown of (B) HSPCs, (C) LT-HSCs, and (D) ST-HSCs isolated from BM at 7 dpi in (G) HSPCs, (H) LT-HSCs, and (I) ST-HSCs, n = 5–7 mice/group. (J and K) (J) Graphic for generation of MethoCult assays, and (K) number of colonies per plate of either WT or Il18r −/− origin that differentiated from mixed chimera BM in MethoCult media. ∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗ p < 0.0001, ∗∗∗∗ p < 0.00001 (see Figure S3 ).

Article Snippet: Mice were administered 75 μg/mouse of anti-IL-18Rα antibodies (clone AF856; R&D Systems) or IgG2 isotype control antibody via i.p. injection on days 4 and 6 post-infection.

Techniques: Irradiation, Isolation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-18 receptor beta-induced changes in the presentation of IL-18 binding sites affect ligand binding and signal transduction.

doi: 10.4049/jimmunol.170.11.5571

Figure Lengend Snippet: FIGURE 1. Expression of IL-18R on L428 and KG-1 cells. IL-18R and IL-18R expressions were analyzed on L428, KG-1, and TNF-- primed KG-1 (pretreated with 10 ng/ml TNF- for 24 h) cells by flow cytometry. The y-axis shows the cell counts, and the x-axis is the signal intensity of PE-labeled anti-IL-18R and FITC-labeled rabbit anti-mouse IgG detecting mouse anti-hIL-18R.

Article Snippet: IL-18R expression was also assessed by flow cytometry using a mouse anti-human IL-18R mAb (R&D Systems, Minneapolis, MN) for cell staining, followed by a secondary FITC-labeled rabbit anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA) for detection.

Techniques: Expressing, Cytometry, Labeling

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-18 receptor beta-induced changes in the presentation of IL-18 binding sites affect ligand binding and signal transduction.

doi: 10.4049/jimmunol.170.11.5571

Figure Lengend Snippet: FIGURE 2. Inhibition of IL-18R binding by anti-IL-18 mAbs and IL- 18BP. Inhibition of binding of 125I-labeled rhIL-18 to L428 (A) or TNF- -primed KG-1 (B) cells by anti-IL-18 mAbs and IL-18BP was determined as described in Materials and Methods. Data are presented as the percent inhibition vs increasing concentrations of IL-18 Abs or IL-18BP.

Article Snippet: IL-18R expression was also assessed by flow cytometry using a mouse anti-human IL-18R mAb (R&D Systems, Minneapolis, MN) for cell staining, followed by a secondary FITC-labeled rabbit anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA) for detection.

Techniques: Inhibition, Binding Assay, Labeling

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-18 receptor beta-induced changes in the presentation of IL-18 binding sites affect ligand binding and signal transduction.

doi: 10.4049/jimmunol.170.11.5571

Figure Lengend Snippet: FIGURE 8. Diagram of IL-18 binding model on L428 and KG-1 cells. The left half of the diagram represents 125-2H inhibition of IL-18 binding to L428 cells expressing only IL-18R. The white circle within the IL-18 molecule represents binding epitope of 125-2H. Since 125-2H binding epitope was directly involved in receptor binding, the interaction of IL-18 with the receptor was abolished in the presence of 125-2H. In primed KG-1 cells, shown on the right half of the picture, the presence of IL-18R may induce a conformational change in IL-18R, resulting in displacement of 125-2H epitope out of the binding pocket. Therefore, the interaction of IL-18 with the receptor complex was not inhibited by 125-2H. However 125-2H epitope might be involved in interacting with IL-18R, which could be critical for signal transduction. As a result, 125-2H can inhibit IL-18-induced IFN- production by KG-1 cells.

Article Snippet: IL-18R expression was also assessed by flow cytometry using a mouse anti-human IL-18R mAb (R&D Systems, Minneapolis, MN) for cell staining, followed by a secondary FITC-labeled rabbit anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA) for detection.

Techniques: Binding Assay, Inhibition, Expressing, Transduction