cxcl 1  (R&D Systems)

Journal: Scientific Reports

Article Title: Peritoneal neutrophil extracellular traps contribute to septic AKI via peritoneal IL-17A and distant organ CXCL-1/ CXCL-2 pathway in abdominal sepsis

doi: 10.1038/s41598-025-34770-1

Figure Lengend Snippet: Knockout of Il-17a attenuated AKI and neutrophil infiltration into kidney and lung, and inhibited CXCL-1 and -2 production in kidney and lung at 18 h after CLP. ( A ) Plasma BUN levels in WT or Il-17a KO mice at 18 h after sham (n = 6 per group) or CLP (n = 10 per group) surgery. ( B ) Tubular damage score in kidney cortex of WT or Il-17a KO mice at 18 h after sham (n = 5 per group) or CLP surgery (n = 8 per group). ( C ) Number of neutrophils in kidney of WT or Il-17a KO mice at 18 h after sham (n = 5 per group) or CLP surgery (n = 8 per group) using naphthol AS-D chloroacetate esterase staining. Neutrophils were counted in × 400 fields and averaged per mouse. ( D ) Number of neutrophils in lung of WT or Il-17a KO mice at 18 h after sham (n = 5 per group) or CLP surgery (n = 9 per group) using naphthol AS-D chloroacetate esterase staining. Neutrophils were counted in × 400 fields and averaged per mouse. ( E , F ) CXCL-1 and -2 concentration in kidney ( E ) and lung ( F ) of WT or Il-17a KO mice at18 h after sham (n = 5 per group) or CLP (n = 8–10 per group) surgery. The data sets were analyzed by one-way ANOVA, followed by Tukey’s multiple comparisons test. Values represent the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: 5 × 10 4 cells were transferred to a poly-L-lysine-coated coverslip (Corning) in 24 well plates, stimulated with 20 ng/ml mouse recombinant IL-17A, CXCl-1, -2 protein (R&D Systems), or 100 nM of phorbol myristate acetate (PMA) for 3 h in a humidified incubator (37 °C, 5% CO 2 ) .

Techniques: Knock-Out, Clinical Proteomics, Staining, Concentration Assay

Journal: Scientific Reports

Article Title: Peritoneal neutrophil extracellular traps contribute to septic AKI via peritoneal IL-17A and distant organ CXCL-1/ CXCL-2 pathway in abdominal sepsis

doi: 10.1038/s41598-025-34770-1

Figure Lengend Snippet: Knockout of Pad4 inhibited CXCL-1 and -2 production in kidney and lung and IL-17A production in peritoneal cavity and plasma at 18 h after CLP. (A-B) CXCL-1 and -2 concentration in kidney ( A ) and lung ( B ) of WT or Pad4 KO mice at18 h after sham (n = 5 per group) or CLP (n = 7 per group) surgery. ( C , D ) IL-17A concentration in PLF ( C ) and plasma ( D ) at 18 h after sham (n = 5 per group) or CLP surgery (n = 7 per group). The data sets were analyzed by one-way ANOVA, followed by Tukey’s multiple comparisons test. Values represent the means ± SEM. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: 5 × 10 4 cells were transferred to a poly-L-lysine-coated coverslip (Corning) in 24 well plates, stimulated with 20 ng/ml mouse recombinant IL-17A, CXCl-1, -2 protein (R&D Systems), or 100 nM of phorbol myristate acetate (PMA) for 3 h in a humidified incubator (37 °C, 5% CO 2 ) .

Techniques: Knock-Out, Clinical Proteomics, Concentration Assay

Journal: Scientific Reports

Article Title: Peritoneal neutrophil extracellular traps contribute to septic AKI via peritoneal IL-17A and distant organ CXCL-1/ CXCL-2 pathway in abdominal sepsis

doi: 10.1038/s41598-025-34770-1

Figure Lengend Snippet: Knockout of Il-17a suppressed CXCL-1 and -2 content in PLF and plasma and H3Cit levels in PLF at 18 h after CLP. ( A , B ) CXCL-1 and -2 concentration in PLF ( A ) and plasma ( B ) of WT or Il-17a KO mice at 18 h after sham (n = 5) or CLP (n = 8) surgery. ( C ) H3Cit levels in PLF of WT or Il-17a KO mice at 18 h after sham (n = 5) or CLP (n = 7–8) surgery. The data sets were analyzed by one-way ANOVA, followed by Tukey’s multiple comparisons test. Values represent the means ± SEM. * P < 0.05, *** P < 0.001, **** P < 0.0001.

Article Snippet: 5 × 10 4 cells were transferred to a poly-L-lysine-coated coverslip (Corning) in 24 well plates, stimulated with 20 ng/ml mouse recombinant IL-17A, CXCl-1, -2 protein (R&D Systems), or 100 nM of phorbol myristate acetate (PMA) for 3 h in a humidified incubator (37 °C, 5% CO 2 ) .

Techniques: Knock-Out, Clinical Proteomics, Concentration Assay

Journal: Scientific Reports

Article Title: Peritoneal neutrophil extracellular traps contribute to septic AKI via peritoneal IL-17A and distant organ CXCL-1/ CXCL-2 pathway in abdominal sepsis

doi: 10.1038/s41598-025-34770-1

Figure Lengend Snippet: Knockout of Il-17a suppressed NET formation in PLF cells ex vivo at 18 h after CLP. Ex vivo stimulation with recombinant IL-17A, CXCL-1, or -2 promoted NET formation in PLF cells. ( A ) Representative images of PLF cells from WT or Il-17a KO mice at 18 h after CLP. PLF cells were applied to a 24 well plate without additional stimulation and incubated at 37 °C for 2 h. These cells were stained with SYTOX green (green) and Hoechst (blue). ( B ) NET extension in PLF cells from WT (n = 10) or Il-17a KO (n = 10) was measured as the SYTOX green-positive area divided by the number of cells detected by Hoechst. ( C ) Representative images of NET formation in PLF cells from WT or Il-17a KO mice at 18 h after CLP. PLF cells were placed on poly-L-lysine coated cover slips without additional stimulation and incubated at 37 °C for 3 h. These cells were stained for H3Cit (Green) and Hoechst (Red pseudocolor). Original magnification, × 400. ( D ) Summary analysis of NET formation in PLF cells of WT (n = 9) or Il-17a KO mice (n = 8) at 18 h after CLP. Percentage of cells with NET formation (upper) and NET extension (lower) in PLF cells were calculated. ( E ) Representative images of NET formation in PLF cells from WT mice at 3 h after CLP. PLF cells were stimulated with recombinant IL-17A, CXCL-1, or -2 at 37 °C for 3 h. These cells were stained for H3Cit (Green) and Hoechst (Red pseudocolor). Original magnification, × 400. ( F ) Summary analysis of NET formation in PLF cells collected 3 h after CLP stimulated with recombinant IL-17A, CXCL-1, -2, or PMA for 3 h (30 fields from 2–3 coverslips per group). Percentage of cells with NET formation (left) and NET extension (right) in PLF cells were calculated. The data sets were analyzed by unpaired t-test ( B and D ) or one-way ANOVA, followed by Tukey’s multiple comparisons test ( F ). Values represent the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Scale bars = 300 µm ( A ) and 20 µm ( C and E ).

Article Snippet: 5 × 10 4 cells were transferred to a poly-L-lysine-coated coverslip (Corning) in 24 well plates, stimulated with 20 ng/ml mouse recombinant IL-17A, CXCl-1, -2 protein (R&D Systems), or 100 nM of phorbol myristate acetate (PMA) for 3 h in a humidified incubator (37 °C, 5% CO 2 ) .

Techniques: Knock-Out, Ex Vivo, Recombinant, Incubation, Staining

Journal: Scientific Reports

Article Title: Peritoneal neutrophil extracellular traps contribute to septic AKI via peritoneal IL-17A and distant organ CXCL-1/ CXCL-2 pathway in abdominal sepsis

doi: 10.1038/s41598-025-34770-1

Figure Lengend Snippet: Schematic presentation of the pathway from NET formation and IL-17A production in the peritoneal cavity to remote organ injuries in the CLP model. CLP upregulated neutrophil accumulation and PAD4 mediated NET formation in the peritoneal cavity (Fig. ). CLP increased IL-17A production in PLF and plasma, which was significantly decreased by Pad4 knockout (Fig. C and D). Knockout of Il-17a significantly decreased CXCL-1 and -2 production in PLF after CLP (Fig. A). Knockout of Il-17a decreased NET formation in peritoneal cavity after CLP (Figs. C and A–D). Recombinant IL-17A, CXCL-1 and -2 upregulated NET formation in PLF cells collected from WT mice at 3 h after CLP ex vivo (Fig. E and F). Pad4 KO as well as Il-17a KO significantly decreased CXCL-1 and -2 production in kidney and lung, neutrophil infiltration into kidney and lung, and AKI after CLP (Figs. and ). Pad4 KO significantly improved survival after CLP (Fig. A). Adoptive transfer of WT neutrophil into Pad4 KO mice restored IL-17A production in PLF and plasma, CXCL-1 and -2 production in kidney and lung, neutrophil infiltration into kidney and lung, and AKI (Fig. ).

Article Snippet: 5 × 10 4 cells were transferred to a poly-L-lysine-coated coverslip (Corning) in 24 well plates, stimulated with 20 ng/ml mouse recombinant IL-17A, CXCl-1, -2 protein (R&D Systems), or 100 nM of phorbol myristate acetate (PMA) for 3 h in a humidified incubator (37 °C, 5% CO 2 ) .

Techniques: Clinical Proteomics, Knock-Out, Recombinant, Ex Vivo, Adoptive Transfer Assay

Journal: Scientific Reports

Article Title: Peritoneal neutrophil extracellular traps contribute to septic AKI via peritoneal IL-17A and distant organ CXCL-1/ CXCL-2 pathway in abdominal sepsis

doi: 10.1038/s41598-025-34770-1

Figure Lengend Snippet: Intraperitoneal adoptive transfer of WT neutrophils counteracted the attenuation of septic AKI, neutrophil infiltration into kidney, IL-17A production in PLF and plasma, CXCL-1/CXCL-2 production in kidney and lung caused by knockout of Pad4 . ( A ) Plasma BUN levels in Pad4 KO mice at 18 h after CLP surgery injected with WT (n = 12) or Pad4 KO (n = 7) neutrophils, or vehicle (n = 5). ( B ) Tubular damage score in kidney cortex in Pad4 KO mice at 18 h after CLP surgery injected with WT (n = 12) or Pad4 KO (n = 7) neutrophils, or vehicle (n = 5). ( C ) Number of neutrophils in kidney of Pad4 KO mice at18 h after CLP surgery injected with WT (n = 12) or Pad4 KO (n = 7) neutrophils, or vehicle (n = 5) using naphthol AS-D chloroacetate esterase staining. Neutrophils were counted in × 400 fields and averaged per mouse. ( D , E ) CXCL-1 and -2 concentration in kidney ( D ) and lung ( E ) of Pad4 KO mice at 18 h after CLP surgery injected with WT (n = 9) or Pad4 KO (n = 7) neutrophils, or vehicle (n = 5). ( F , G ) IL-17A concentration in ( F ) PLF and ( G ) plasma of Pad4 KO mice at 18 h after CLP surgery injected with WT (n = 12) or Pad4 KO (n = 7) neutrophils, or vehicle (n = 5). The data sets were analyzed by one-way ANOVA, followed by Tukey’s multiple comparisons test. Values represent the means ± SEM. ns: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars = 20 µm.

Article Snippet: 5 × 10 4 cells were transferred to a poly-L-lysine-coated coverslip (Corning) in 24 well plates, stimulated with 20 ng/ml mouse recombinant IL-17A, CXCl-1, -2 protein (R&D Systems), or 100 nM of phorbol myristate acetate (PMA) for 3 h in a humidified incubator (37 °C, 5% CO 2 ) .

Techniques: Adoptive Transfer Assay, Clinical Proteomics, Knock-Out, Injection, Staining, Concentration Assay