fc fused human il 18 binding protein isoform  (R&D Systems)

Journal: eBioMedicine

Article Title: Danger signals released during cold ischemia storage activate NLRP3 inflammasome in myeloid cells and influence early allograft function in liver transplantation

doi: 10.1016/j.ebiom.2022.104419

Figure Lengend Snippet: Quantification of several DAMPs and different proteins detected in OPS collected after cold ischemia static storage.

Article Snippet: Proteinase K was from Roche (Basel, Switzerland), DNase I from Qiagen (Hilden, Germany), Uricase from Worthington (Lakewood, USA), anti-HMGB1 (Cat# 651,401; RRID: AB_10945159 ) from Biolegend (San Diego, USA), and recombinant human IL-18 binding protein (IL-18BP) from GeneScript (Piscataway, USA).

Techniques: Concentration Assay

Journal: The Journal of Neuroscience

Article Title: Delayed Activation of Spinal Microglia Contributes to the Maintenance of Bone Cancer Pain in Female Wistar Rats via P2X7 Receptor and IL-18

doi: 10.1523/JNEUROSCI.5250-14.2015

Figure Lengend Snippet: P2X7R mediates bone cancer-induced IL-18 upregulation via p38 MAPK in the spinal dorsal horn. A, Western blot analysis reveals significant upregulation of p-p38 levels on PTDs 14 and 21. **p < 0.01 versus sham rats. B, Knockdown P2X7R significantly reduces bone cancer-induced activation of p38 on PTD 14. **p < 0.01 versus control siRNA (Cntr.). C, Western blot analysis reveals significant upregulation of IL-18 levels on PTDs 14 and 21. **p < 0.05 versus sham rats. D, Knockdown P2X7R significantly suppressed bone cancer-induced upregulation of IL-18 on PTD 14. **p < 0.01 versus control siRNA. E, Selective p38 inhibitor SB239063 (SB; 10 μg, i.t.) markedly suppressed the bone cancer-induced increase in IL-18 levels on PTD 14. F, G, Intrathecal injection of SB239063 or IL-18BP (an endogenous inhibitor of IL-18 activity; 1 μg, i.t.) reversed bone cancer-induced mechanical allodynia (F) and hyperalgesia (G) on PTD 14. *p < 0.05, **p < 0.01 versus vehicle controls (PBS or DMSO).

Article Snippet: The inhibitor of microglial activation minocycline (Sigma-Aldrich), astrocyte metabolic inhibitor fluorocitrate (Sigma-Aldrich), P2X7 antagonist brilliant blue G (BBG; Sigma-Aldrich), recombinant human IL-18 binding protein [IL-18BP (an inhibitor of IL-18 activity); R&D Systems], and recombinant rat IL-18 (R&D Systems) were dissolved and diluted in sterile PBS.

Techniques: Western Blot, Activation Assay, Injection, Activity Assay

Journal: The Journal of Neuroscience

Article Title: Delayed Activation of Spinal Microglia Contributes to the Maintenance of Bone Cancer Pain in Female Wistar Rats via P2X7 Receptor and IL-18

doi: 10.1523/JNEUROSCI.5250-14.2015

Figure Lengend Snippet: Colocalization of P2X7R/p-p38 and IL-18 in spinal microglia. A, Double immunofluorescence of p-p38 with IL-18-IR, CD11b-IR, NeuN-, IR, and GFAP-IR on the ipsilateral spinal dorsal horn on PTD 14. Note the heavy colocalization of p-p38-IR with IL-18-IR and CD11b-IR. B, Double immunofluorescence shows colocalization of IL-18 with P2X7R and IBA-1 on the ipsilateral spinal dorsal horn on PTD 14.

Article Snippet: The inhibitor of microglial activation minocycline (Sigma-Aldrich), astrocyte metabolic inhibitor fluorocitrate (Sigma-Aldrich), P2X7 antagonist brilliant blue G (BBG; Sigma-Aldrich), recombinant human IL-18 binding protein [IL-18BP (an inhibitor of IL-18 activity); R&D Systems], and recombinant rat IL-18 (R&D Systems) were dissolved and diluted in sterile PBS.

Techniques: Immunofluorescence

Journal: The Journal of Neuroscience

Article Title: Delayed Activation of Spinal Microglia Contributes to the Maintenance of Bone Cancer Pain in Female Wistar Rats via P2X7 Receptor and IL-18

doi: 10.1523/JNEUROSCI.5250-14.2015

Figure Lengend Snippet: P2X7R/IL-18 signaling regulates neuronal activity in the spinal dorsal horn. A, Histograms show typical responses of a WDR neuron to brush and pinch delivered to the receptive fields in a PTD 14 rat. Oscilloscope recording shows a single sweep (inset). B, C, Histograms show the typical responses of WDR neurons to von Frey filament stimuli delivered to the receptive fields in a naive rat (B) and a PTD 14 rat (C). D, E, The average number of discharges evoked by brush and pinch (D) and von Frey hairs (E) in PTD 14 rats is significantly higher than in sham rats. **p < 0.01 versus sham controls. F, G, Blockade of P2X7R by BBG or the inhibition of IL-18 by IL-18BP significantly suppresses the evoked responses of WDR neurons by von Frey hairs (F), and brush and pinch (G) stimuli in PTD 14 rats. *p < 0.05, **p < 0.01 versus vehicle (Veh) control. H, Histograms show the facilitatory effect of IL-18 on responses of WDR neurons to von Frey stimuli delivered to the receptive fields in a naive rats. I, J, Intrathecal injection of exogenous IL-18 (3 μg) enhances the evoked response of WDR neurons by von Frey hairs (I), and brush and pinch (J) stimuli in naive rats. K, Intrathecal injection of IL-18 of different doses (0.03, 0.3, and 3 μg) induces allodynia in naive rats. L, Pretreatment of fluorocitrate (FC; 0.8 μg = 1 nmol, i.t., 30 min before IL-18 injection) did not block IL-18-induced allodynia. *p < 0.05, **p < 0.01 versus vehicle control.

Article Snippet: The inhibitor of microglial activation minocycline (Sigma-Aldrich), astrocyte metabolic inhibitor fluorocitrate (Sigma-Aldrich), P2X7 antagonist brilliant blue G (BBG; Sigma-Aldrich), recombinant human IL-18 binding protein [IL-18BP (an inhibitor of IL-18 activity); R&D Systems], and recombinant rat IL-18 (R&D Systems) were dissolved and diluted in sterile PBS.

Techniques: Activity Assay, Inhibition, Injection, Blocking Assay

Journal: The Journal of Neuroscience

Article Title: Delayed Activation of Spinal Microglia Contributes to the Maintenance of Bone Cancer Pain in Female Wistar Rats via P2X7 Receptor and IL-18

doi: 10.1523/JNEUROSCI.5250-14.2015

Figure Lengend Snippet: IL-18 modulates mEPSCs in the superficial dorsal horn neurons. A, Patch-clamp recording of mEPSC shows an increase in the frequency of mEPSCs after perfusion of IL-18 (10 ng/ml). B, C, Corresponding cumulative distributions. D, Quantification of mEPSC frequency. E, Quantification of mEPSC amplitudes. **p < 0.01 versus control. F, mEPSC traces show an increase in the frequency of mEPSCs after the perfusion of IL-18 in the presence of fluorocitrate (FC; 10 μm). G, H, Corresponding cumulative distributions. I, Ratio of the frequency and amplitude of mEPSCs following treatment with IL-18, FC, and FC plus IL-18. Dashed line indicates baseline. Rats were pretreated with FC for 30 min before the application of IL-18. *p < 0.05, **p < 0.01 versus control; #p < 0.05, ##p < 0.01 versus FC alone.

Article Snippet: The inhibitor of microglial activation minocycline (Sigma-Aldrich), astrocyte metabolic inhibitor fluorocitrate (Sigma-Aldrich), P2X7 antagonist brilliant blue G (BBG; Sigma-Aldrich), recombinant human IL-18 binding protein [IL-18BP (an inhibitor of IL-18 activity); R&D Systems], and recombinant rat IL-18 (R&D Systems) were dissolved and diluted in sterile PBS.

Techniques: Patch Clamp

Journal: The Journal of Neuroscience

Article Title: Delayed Activation of Spinal Microglia Contributes to the Maintenance of Bone Cancer Pain in Female Wistar Rats via P2X7 Receptor and IL-18

doi: 10.1523/JNEUROSCI.5250-14.2015

Figure Lengend Snippet: Schematic illustration of neuron–glia and glia–glia interactions in the spinal cord dorsal horn in bone cancer pain. Bone cancer-induced hyperexcitability of primary sensory neurons (peripheral sensitization) causes excessive release of neurotransmitters or neuromodulators from central afferent terminals to activate adjacent glia and postsynaptic neurons in the spinal dorsal horn. P2X7R is upregulated in microglia and activated by ATP, which is produced by activated astrocytes, as well as by primary afferent terminals and spinal cord neurons. Upon activation, microglia synthesize and release IL-18 via phosphorylation of p38 MAPK, leading to enhanced excitatory synaptic transmission and neuronal hyperactivity in the dorsal horn (central sensitization). As a result of this IL-18-mediated neuromodulation in the spinal cord pain circuit, pain sensitivity is enhanced. Solid lines indicate the pathways and mechanisms demonstrated in this study. Dashed lines indicate other possible pathways and mechanisms.

Article Snippet: The inhibitor of microglial activation minocycline (Sigma-Aldrich), astrocyte metabolic inhibitor fluorocitrate (Sigma-Aldrich), P2X7 antagonist brilliant blue G (BBG; Sigma-Aldrich), recombinant human IL-18 binding protein [IL-18BP (an inhibitor of IL-18 activity); R&D Systems], and recombinant rat IL-18 (R&D Systems) were dissolved and diluted in sterile PBS.

Techniques: Produced, Activation Assay, Transmission Assay