il 8  (R&D Systems)

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Concentration-dependent change in the humoral inflammatory response following incubation with Escherichia coli ( E. coli ) in the ex vivo whole blood model. a Absolute plasma concentrations of IL-6, IL-8, and MMP9 determined by enzyme-linked immunosorbent assay. b Normalized values and EC 50 curve fit by BuC=0% and 50 000 CFU/ml E. coli= 100%, respectively, for IL-6, IL-8, and MMP9 as indicated by EC 50 (%) on the respective Y-axis. BuC indicates buffer control after 60 min incubation; numbers on the X-axis indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation; LPS indicates lipopolysaccharide (LPS) 100 ng/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, comparing all shown concentrations of E. coli bacteria and 100 ng/ml LPS with BuC. P -values are indicated above the respective data points. ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Concentration Assay, Incubation, Ex Vivo, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Bacteria

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Diagnostic performance for the detection of bacteremia, analyzing the neutrophil phenotype by determining the median fluorescence intensity (MFI) and the cellular response capacity (CRC) in comparison with traditional markers of humoral inflammation (IL-6, IL-8, MMP9). a Comparison of receiver operating characteristic (ROC) at 10,000 CFU/ml Escherichia coli ( E. coli ) with the respective 95% confidence interval (CI) and P -value, and half-maximal effective concentration (EC 50 ) as a function of the E. coli concentration. b Detailed comparison of the EC 50 as a function of the E. coli concentration. c Exemplary comparison of EC 50 curve fit after normalization as indicated by EC 50 (%) on the respective Y-axis to BuC=100% and 50 000 CFU/ml E. coli =0% for the humoral marker IL-6 (the IL-6 values were multiplied by −1 before EC 50 calculation to facilitate comparability with the CRC) and the change in neutrophil phenotype represented by CD11b CRC. BuC indicates buffer control after 60 min incubation; numbers on the X-axis of c indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, evaluating the EC 50 of IL-8, MMP9, the MFI, and CRC of CD10, CD11b, and CD62L in comparison to the EC 50 of IL-6. P -values are indicated above the respective data points. ⁎ P <0.05. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Diagnostic Assay, Fluorescence, Comparison, Concentration Assay, Marker, Control, Incubation, Bacteria

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Clinical specifications and parameters over all time points of the sepsis cohort. a Suspected infection cause of sepsis. b Distribution of the individual score points of the Sequential Organ Failure Assessment (SOFA) score. c Total SOFA score. d-h Traditional and humoral markers of inflammation: leukocytes and neutrophil-lymphocyte ratio ( d ), C-reactive protein (CRP) and procalcitonin (PCT) ( e ), interleukin-6 (IL-6) and interleukin-8 (IL-8) ( f ), serum amyloid A (SAA) and calprotectin ( g ), matrix metallopeptidase 9 (MMP9) and myeloperoxidase (MPO) ( h ). Values are shown as median and interquartile range. n =14. CNS. Central nervous system; HV. Healthy volunteers.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Infection

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) Cell viability was assessed with the MTT assay and data shown are means ± SD (n = 3). ( b ) TNFα, IL-6 and IL-8 expression in response to B[a]P exposure. ( c ) Representative Western blot images showing protein expression. All data were obtained from three independent experiments. Statistical p values were determined by a one-way ANOVA, followed by a Tukey post-hoc test comparing the B[a]P treatments to the DMSO control; * p < 0.05; ¥, p < 0.01; NS, no significance.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: MTT Assay, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: Cells were treated with various concentrations of PM2.5 for 24 hr. ( a ) Cell viability was assessed with MTT assay. ( b ) IL-6 and IL-8 from the basolateral sides. ( c ) Representative Western blot images showing CFTR, β2AR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), NF-κB p65, and TRPC6 expression. All data are presented as the mean ± SD (n = 4). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; *, p < 0.05; ¥, p < 0.01; NS, no significance.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: MTT Assay, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) IL-6 and IL-8 from the basolateral sides of the ALI were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n = 4). Statistical p -values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the all treatments to the DMSO control; ¥, p < 0.05; *, p < 0.01. ( b ) Representative Western blot images showing CFTR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), and NF-κB p65 expression in treated polarized 16HBE14o-cells in ( a ). All data were obtained from three independent experiments.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ALI cells were treated with DMSO, 100 ng/mL PM 2.5, 10 µM LY 294002 ± PM 2.5, or 10 µM LY 303511 ± PM 2.5 for 24 hrs. ( a ) IL-6 and IL-8 from the basolateral sides were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n=3). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing all treatments to the DMSO control, or PM 2.5 to PM 2.5 ± LY 294002 or LY 303511; *, p < 0.01. ( b ) Representative Western blot images showing CFTR and β-actin expression. β-actin was used to ensure equal loading of protein. All data were obtained from three independent experiments, and each experiment was analyzed in duplication.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) The lung parenchyma from two ferrets were sliced into several sections as shown. Arrows indicate the tracheobronchial tubes. All tissue sections were incubating with DMSO (control), 10 µM B[a]P, or 100 ng/mL PM 2.5 in Pneumacult-EX Plus Basal media for 24 hrs. ( b ) IL 6 secretion was determined using the ferret ELISA kit. Data represent the mean of 6 independent sections. Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; ¥, p < 0.05 (n = 6). ( c ) Representative western blotting images show reduction of CFTR, and β2AR and elevation of TRPC6 expression in explants under these treatments.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing