Journal: Cancer immunology research

Article Title: NET-Triggered NLRP3 Activation and IL18 Release Drive Oxaliplatin-Induced Peripheral Neuropathy.

doi: 10.1158/2326-6066.CIR-22-0197

Figure Lengend Snippet: Figure 5. NLRP3 activation and subsequent IL18 release contribute to oxaliplatin-induced mechanical hyperalgesia. A, The level of ROS in the DRG at day 14 after the initiation of L-OHP treatment was assessed by DCFH-DA staining (n ¼ 8). B and C, The level of ROS in BMDMs stimulated with L-OHP (1, 5, or 10 mmol/L) for 1 hour was assessed by calculating the ratio of DCFH-DA–positive cells to 10,000 cells through flow cytometry. BMDMs stimulated with rotenone (10 mmol/L, 6 hours) were used as a positive control (n ¼ 4). D–K, BMDMs were stimulated with NETs for 3 hours and then stimulated with L-OHP (10 mmol/L) or left unstimulated for 1 hour. LPS (1 mg/mL)-primed BMDMs were treated with ATP (1.5 mmol/L) as a positive control (n ¼ 3). Western blot was used to assess the activation of NLRP3 inflammasome–related proteins [NLRP3, Pro-IL1b, Pro-caspase-1, Caspase-1 (p20), IL1b] in BMDMs treated with NETs and/or L-OHP in both culture supernatants [caspase-1 (p20), IL1b] and cell lysates (NLRP3, Pro-IL1b, Pro-caspase-1; D-I). IL1b or IL18 in supernatants were analyzed by ELISA (J and K). L–Q, WT and Pad4/

Article Snippet: To verify the ability of IL18BP to inhibit IL18 signaling in vivo, recombinant mouse IL18 (HY-P73181, MedChemExpress; 20 ng per mouse, i.p.) was administered toNlrp3 / mice for 7 consecutive days.

Techniques: Activation Assay, Staining, Cytometry, Positive Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cancer immunology research

Article Title: NET-Triggered NLRP3 Activation and IL18 Release Drive Oxaliplatin-Induced Peripheral Neuropathy.

doi: 10.1158/2326-6066.CIR-22-0197

Figure Lengend Snippet: Figure 6. NETs and IL18 are detected in the plasma of patients after chemotherapy. A and B, The level of H3Cit and cfDNA, as measured by H3Cit ELISA kit and Quant-iTPico green dsDNA assay, respectively, in plasma from healthy volunteers and cancer patients who had or had not received chemotherapy (n ¼ 10). C and D, The levels of IL18 and IL1b in the same plasma samples used in A–B were measured by ELISA (n ¼ 10). E, Correlations between IL18 and H3Cit were analyzed by linear regression analysis (R2 ¼ 0.6993, P ¼ 0.0026). F, Correlations between IL18 and cfDNA were analyzed by linear regression analysis (R2 ¼ 0.5209, P ¼ 0.0184). G–I, Correlations between VAS and H3Cit, cfDNA or IL18 were analyzed by linear regression analysis (R2 ¼ 0.7673, P ¼ 0.0009; R2 ¼ 0.7107, P ¼ 0.0022; R2 ¼ 0.5148, P ¼ 0.0195, respectively). , P < 0.05; , P < 0.001. Data are shown as mean SEM.

Article Snippet: To verify the ability of IL18BP to inhibit IL18 signaling in vivo, recombinant mouse IL18 (HY-P73181, MedChemExpress; 20 ng per mouse, i.p.) was administered toNlrp3 / mice for 7 consecutive days.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, dsDNA Assay