Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: DAMP S100A8 along with NLRP3 inflammasome-dependent pyroptosis is positively related to the progression of liver fibrosis. ( A ) Representative IHC images for S100A8 and S100A9 in liver sections from liver fibrosis patients and HCs. ( B , C ) ELISA analyses for serum levels of S100A8 and S100A9 in liver fibrosis patients and HCs. ( D ) Comparison of serum S100A8 and S100A9 levels in liver fibrosis patients with different phases. ( E – G ) Distribution of serum GSDMD ( E ), IL-1β ( F ), and IL-18 ( G ) levels in liver fibrosis patients with different phases (F0–4). ( H – J ) Correlation between serum S100A8 levels and GSDMD ( H ), IL-1β ( I ) or IL-18 ( J ) levels in liver fibrosis patients. ( K ) Representative mouse liver morphology and staining with H&E and Sirius Red. ( L – O ) IHC staining of mouse liver sections for NLRP3, S100A8, and S100A9. Black scale bar: 100 µm; Red scale bar: 50 µm. ELISA analyses for serum levels of S100A8 ( L ), GSDMD ( M ), IL-1β ( n ), and IL-18 ( O ) in 4-, 6-, and 8 week-mouse models of liver fibrosis. *** p < 0.001.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Staining, Immunohistochemistry

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: S100A8-mediated NLRP3 inflammasome-dependent pyroptotic macrophage death amplify the activation of human hepatic stellate cells. ( A – C ) The qRT–PCR analysis for the mRNA levels of NLRP3, pro-IL-1β, and pro-IL-18 in THP-1 macrophages treated with 0, 2, 5 or 10 µg/mL rhS100A8 or 5 µg/mL GST for 24 h. ( D ) The protein levels of NLRP3, GSDMD, GSDMD P30, pro-IL-1β, mature IL-1β, and cleaved caspase-1 were detected by Western blot in THP-1 macrophages treated with 5 µg/mL GST or rhS100A8. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). ( E ) PI and active caspase-1 double staining of pyroptotic cell death by flow cytometry in THP-1 macrophages treated with 5 µg/mL GST or rhS100A8. ( F – I ) Western blot analysis ( F ) and qRT-PCR analysis ( G–I ) of COL1A1, α-SMA, and TGF-β in LX-2 cells exposed to CM from THP-1 macrophages that were treated with 0, 2, 5 or 10 µg/mL of rhS100A8 or 5 µg/mL GST. ( J ) Western blot analysis of COL1A1, α-SMA, and TGF-β in LX-2 cells exposed to CM from THP-1 macrophages that were treated with 5 µg/mL of rhS100A8 with or without 1 h of MCC950 pretreatment. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Activation Assay, Quantitative RT-PCR, Western Blot, Expressing, Double Staining, Flow Cytometry

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: TLR4/NF-κB signaling cascade and ROS abundance are responsible for S100A8-induced NLRP3 inflammasome-dependent pyroptotic death in macrophages. ( A ) Western blot analysis of p65, p-p65, IKKα, and p-IKKα expression in THP-1 macrophages treated with GST-rhS100A8 or GST for 0, 30, 60 or 120 min. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the 0 min group (right panel). ( B – E ) THP-1 macrophages were exposed to 5 µg/mL rhS100A8 with or without 1 h of BAY 11-7082, TAK-242 or FPS-ZM1 pretreatment. The qRT-PCR analysis was performed to detect the mRNA levels of NLRP3 ( B ), pro-IL-1β ( C ), and pro-IL-18 ( D ). Western blot analysis was used to determine the protein expression of NLRP3, GSDMD, GSDMD P30, pro-IL-1β, mature IL-1β, and cleaved caspase-1 ( E ). The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). ( F ) THP-1 macrophages were pretreated with TAK-242 or FPS-ZM1 for 1 h and then exposed to 5 µg/mL of rhS100A8. Western blot analysis was used to determine the expression of p-p65 and p-IKKα. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). ( G ) Flow cytometry analysis of ROS levels in THP-1 macrophages treated with rhS100A8 for 6 h. ( H ) THP-1 macrophages were exposed to 5 µg/mL of rhS100A8 with or without 1 h of DPI pretreatment. Protein expression levels of NLRP3, GSDMD, GSDMD P30, pro-IL-1β, and mature IL-1β were determined by Western blot. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel); ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Flow Cytometry

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: The potential predictive powers of S100A8, GSDMD, IL-1β, and IL-18 for the occurrence and severity of liver fibrosis. ( A ) ROC curves of serum S100A8, GSDMD, IL-1β, and IL-18 for distinguishing liver fibrosis patients from HCs. ( B ) ROC curve, of serum S100A8, GSDMD, IL-1β, and IL-18 for detecting moderate-to-severe liver fibrosis from no or mild liver fibrosis in liver fibrosis patients. ( C ) A working model illustrating that S100A8-mediated NLRP3 inflammasome-dependent pyroptosis in macrophages facilitates liver fibrosis progression, and that the identified GSDMD may be used as a potential biomarker during liver fibrosis onset and progression.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Biomarker Discovery

Journal: NPJ Vaccines

Article Title: Protective efficacy of the pan-fungal vaccine NXT-2 against vulvovaginal candidiasis in a murine model

doi: 10.1038/s41541-025-01171-4

Figure Lengend Snippet: IL-1β ( A ) and S100A8 ( B ) concentrations were assessed via ELISA. All data represents mean ± SEM. Statistical significance was assessed by using the Mann-Whitney test for IL-1β and S100A8 concentrations. C Tissue damage was assessed by measuring lactate dehydrogenase (LDH) activity in vaginal lavage fluid twenty-eight days post vaccination (28dpv) and five days post challenge (5dpc). Data represents mean ± SEM. Statistical significance was assessed by two-way ANOVA with Tukey correction (* P = 0.0111, *** P = 0.0007, **** P < 0.0001). D Quantification of inflammation in vaginal tissue from sham and NXT-2 immunized mice excised 5 days post challenge. Data represents mean ± SEM. Statistical significance was assessed by unpaired student t -test (**** P < 0.0001). Vaginal tissue excised following immunization and C. albicans challenge was stained with H&E (sham— E , NXT-2— G ) and the neutrophil specific marker anti-mouse Ly6G (sham— F , NXT-2— H ). Tissue sections were scanned at 40× objective magnification using the Aperio AT2 (Leica Biosystems). Image analysis and capture was performed using ImageScope (Leica Biosystems) at 20× digital zoom level.

Article Snippet: S100A8 : Utilizing the Mouse S100A8 ELISA kit (Rockland Immunochemicals), samples were diluted 1:100 and the manufacturer’s instructions were followed.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Activity Assay, Staining, Marker