il 8  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: Anti-inflammatory macrophages limit endothelial inflammation and restrict neutrophil recruitment through Gas6/AXL signaling

doi: 10.64898/2026.10.12.758565

Figure Lengend Snippet: (A) Schematic of the neutrophil extravasation assay. The TNF-α-primed HUVEC lumen is loaded with freshly isolated human neutrophils, a chemoattractant solution containing IL-8 and LTB 4 is added to the top port, and confocal imaging tracks neutrophil extravasation and migration over 8 h. (B) Anti-inflammatory experiment, normalized neutrophil extravasation for anti-inflammatory devices versus their macrophage-free control. *P < 0.05. N = 9 paire(D) n = 2–3. (C) Anti-inflammatory experiment, neutrophil migration distance from the lumen edge. *P < 0.05, **P < 0.01. N = 8–9 paire(D) n = 2–3. (D) Pro-inflammatory experiment, normalized neutrophil extravasation for pro-inflammatory devices versus their macrophage-free control. P > 0.05. N = 10–11 paire(D) n = 2–3. (E) Pro-inflammatory experiment, neutrophil migration distance from the lumen edge. P > 0.05. N = 10–11 paire(D) n = 2–3. The pro-inflammatory and anti-inflammatory experiments were performed independently, each with its own macrophage-free control, and analyzed by within-experiment paired two-sample t-test at each timepoint without multiple-comparison correction. In (B) to (E), the center line is the median, the box spans the interquartile range and the whiskers span the full rang(E) with individual biological replicates overlaid. All devices received inflammatory stimulation.

Article Snippet: Unless stated otherwise, inflammatory stimulation means TNF-α (50 ng/mL; R&D Systems, 210-TA-020/CF) introduced into the lumen for 2 h at 37 °C followed by three EGM-2 washes, together with a chemoattractant solution of IL-8 (10 μM; Miltenyi Biotec, 130-122-353) and LTB 4 (100 nM; Cayman Chemical, 20110) in the top port.

Techniques: Isolation, Confocal, Imaging, Migration, Control, Comparison

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: Chembiochem

Article Title: Functional Heterogeneity of the Autism Spectrum Disorder‐Associated Gut Microbial Ecosystem Revealed by Fecal Lipopolysaccharides and Bacterial Proteomic Profiling

doi: 10.1002/cbic.70557

Figure Lengend Snippet: Effects of fecal LPS on HMC3 microglia cell viability and activation. (A) Bright‐field image of HMC3 cells (40× magnification, scale bar: 20 µm). (B) HMC3 microglia cell viability after 24 h exposure to increasing concentrations (0.1, 1, 10, 100 ng/mL) of the pooled fecal LPS preparations obtained from the ASD and NASD cohorts. Cell viability was expressed as % relative to non‐stimulated (NS) cells and assessed by MTT assay (570 nm). NS cells were also considered as negative control. (C,D) Quantification of (C) IL‐8 and (D) IL‐6 levels following LPSs treatment, measured by DuoSet‐ELISA kit. Three independent experiments ( n = 3) were performed, with each experimental condition tested in triplicate wells within each experiment. Statistical significance was assessed by ordinary one‐way ANOVA followed by Tukey's multiple‐comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus NS; # p < 0.05, ### p < 0.001, #### p < 0.0001, versus LPS.

Article Snippet: Cytokine levels (pg/mL) in HMC3 supernatants were measured using DuoSet ELISA kits (R&D Systems, Minneapolis, MN, USA) for human IL‐8 (#DY208) and IL‐6 detection (#DY206).

Techniques: Activation Assay, MTT Assay, Negative Control, Enzyme-linked Immunosorbent Assay, Comparison