c5a elisa kit mouse duo set  (R&D Systems)

Journal: European Journal of Trauma and Emergency Surgery

Article Title: Multifaceted intestinal defense following experimental blunt abdominal trauma

doi: 10.1007/s00068-026-03145-0

Figure Lengend Snippet: Posttraumatic defensive pathway activation and cellular immune response in the murine jejunum. ( A ) Proteome profiler-based pathway analysis revealed activation of immune and defense-related mechanisms. The 15 pathways with the highest degree of regulation are shown. ( B, C ) Histological quantification in jejunal tissue demonstrated a significant increase of CD3 + T lymphocytes in the trauma group relative to controls. ( D ) Gene expression profiling of S100a8 showed a significant post-traumatic upregulation in traumatized mice compared to sham animals. ( E ) In contrast, no statistically significant alteration was observed in interleukin-6 (Il6) expression. ( F ) Plasma levels of C5a were unaltered by traumatic exposure. ( G ) C5a in bronchoalveolar lavage fluid (BALF) was significantly increased in mice 24 h after AT compared to shams. C : n = 180 per group; D-G : n = 4–5 per group. Statistical analysis was performed using the Mann-Whitney U test, with significance defined as p < 0.05. Data are presented as mean ± standard error of the mean (SEM)

Article Snippet: To measure the concentrations of C5a in plasma and BALF, a sandwich ELISA was performed using the C5a ELISA Kit Mouse Duo Set (R&D Systems, Minneapolis, USA).

Techniques: Activation Assay, Gene Expression, Expressing, Clinical Proteomics, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Reactive oxygen species drive the aberrant immune response to a C. neoformans chitin synthase 3 ( chs3Δ ) mutant

doi: 10.3389/fimmu.2025.1691076

Figure Lengend Snippet: Differential regulation of CXCR2 ligands may drive neutrophil recruitment in the absence of ROS: Male and female C57BL/6 or gp91 phox- mice were inoculated with 10 7 CFUs of HK chs3∆ . BAL was collected from each group at 12 h post-inoculation, as well as a phosphate-buffered saline (PBS) control for each group. (A) LBT4, (B) C5a, (C) CXCL1/KC, and (D) CXCL2/MIP-2α. Data are cumulative from two experiments, with a total of 8 mice per group per time point. Values are means ± standard errors of the means (SEM). (**, P <0.005, ***, P <0.001).

Article Snippet: Cytokines were analyzed using Mouse CXCL1/KC ELISA kit (R&D Systems), Mouse CXCL2/MIP-2α ELISA kit (R&D Systems), and Mouse C5a ELISA kit (R&D Systems).

Techniques: Saline, Control

Journal: eBioMedicine

Article Title: Directed-complement killing of Pseudomonas aeruginosa protects against lethal pneumonia

doi: 10.1016/j.ebiom.2025.105926

Figure Lengend Snippet: Anti-Psl CoMiX bind specifically to P. aeruginosa , but not to other bacterial strains, with CoMiX-FHR1 competing with FH. The binding of anti- P. aeruginosa CoMiX, irrelevant anti- A. fumigatus CoMiX, and irrelevant anti- M. catarrhalis CoMiX to reference strain PAO1 (a) , clinical isolate IT 2 (b) , and Gram-bacterial strain M. catarrhalis (c) was analysed by whole-cell ELISA. Immobilised bacteria (1 × 10 7 CFU/mL) or bacterial lysate (20 μg/mL) were incubated with 10 μg/mL of CoMiX and irrelevant controls. Bound CoMiX was detected using specific antibodies: anti-His for CoMiX-FHR1 and anti-Fc for CoMiX-Fc. CoMiX targeting the second bacterial strain and the fungi were used as negative control. Data are presented as the mean values ± SEM. Results correspond to two-three pooled independent experiments (2–3 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. ∗∗∗∗p < 0.0001. To investigate the competition between FH and CoMiX-FHR1, recombinant FH (20 μg/mL) and increasing concentrations of CoMiX-FHR1 (and CoMiX-Fc as control) were added to PAO1 bacteria. The binding of FH to bacterial cells was detected with the FH-specific monoclonal antibody OX-24 (d) . Data are presented as the mean values ± SEM. Results correspond to two-three pooled independent experiments (3 replicates per experiment). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test: it indicates significant differences for the binding of FH in absence versus presence of CoMiX-FHR1. ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.

Article Snippet: To measure anaphylatoxin C5a concentrations in the BAL and the lungs, we used a C5a mouse ELISA kit (#DY2150, R&D Systems, United States) following the manufacturer's instructions.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Bacteria, Incubation, Negative Control, Recombinant, Control

Journal: eBioMedicine

Article Title: Directed-complement killing of Pseudomonas aeruginosa protects against lethal pneumonia

doi: 10.1016/j.ebiom.2025.105926

Figure Lengend Snippet: Anti-Psl CoMiX enhance C1q deposition (for CoMiX-Fc), C3b opsonisation and the formation of membrane attack complex (C5b9/MAC) on P. aeruginosa . Immobilised bacterial cells of the P. aeruginosa reference strain PAO1 and the clinical isolate IT 2 were incubated with 10 μg/mL of CoMiX, before the addition of either 2% (C3b) or 4% (C1q, C5b9/MAC) of normal human serum (NHS) or heat-inactivated human serum (ΔNHS) in GVB++ at 37 °C for 30 min. Complement deposition at the bacterial surface was measured by ELISA, using (a) an anti-human C1q mAb, (b) an anti-human C3/C3b/iC3b mAb, and (c) an anti-human C5b9 mAb, followed by an HRP-conjugated anti-mouse IgG mAb. Data are presented as the mean values ± SEM. Results correspond to three pooled independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. GFP-expressing PAO1 bacteria were incubated with 10% C5-deficient serum (C3b) or normal human serum (C5b9/MAC) at 37 °C for 1 h, in the presence or absence of 10 μg/mL of CoMiX. Bacteria were then incubated with either a goat anti-human C3b or goat anti-human C5b9 antibody, followed by a secondary AF647-conjugated anti-goat mAb. Once stained, bacteria were mounted onto an agarose gel pad, visualised on a confocal Leica Sp8 microscope (C3b), or on a wide field Axio observer microscope (C5b), and analysed by ImageJ to detect C3 cleavage product (C3b, iC3b, and C3c) deposition (d) or C5b9 deposition (e) . Two to four fields have been acquired for each conditions. Representative images of the deposition are presented here. Green = GFP-expressing PAO1, red = C3b or C5b9 deposition by anti-goat AF647. Scale bar = 5 μm; 10 μm.

Article Snippet: To measure anaphylatoxin C5a concentrations in the BAL and the lungs, we used a C5a mouse ELISA kit (#DY2150, R&D Systems, United States) following the manufacturer's instructions.

Techniques: Membrane, Incubation, Enzyme-linked Immunosorbent Assay, Expressing, Bacteria, Staining, Agarose Gel Electrophoresis, Microscopy

Journal: eBioMedicine

Article Title: Directed-complement killing of Pseudomonas aeruginosa protects against lethal pneumonia

doi: 10.1016/j.ebiom.2025.105926

Figure Lengend Snippet: Therapeutic administration of CoMiX in vivo results in enhanced bacterial clearance through local and systemic complement activation. Mice were infected/treated as described in a, sacrificed at 4 h, 8 h and 16 h p.i. , and lungs, BAL and blood were collected for analysis. (a) Lungs were first perfused for a visual assessment of inflammation. Bacterial load in BAL (b) and lungs (c) were determined after serial dilution and plating on Petri dishes. Total protein was measured by BCA in the BAL (d) . To assess the activation of the complement cascade, the concentration of activated fragments of the mouse complement protein C3 was determined by ELISA, systemically in the serum (e) and locally in the BAL (f) . Concentration of local C3 activated fragments was correlated with the BAL bacterial load (g) . The concentration of mouse complement protein C5a was determined by ELISA locally in the BAL (h) and the lungs (i) . All data are shown as individual values and quoted as the mean values ± SEM and the results correspond to one experiment per time-point ( n = 7–8 mice per group). Unless otherwise stated, all statistical analyses were performed using Kruskal–Wallis test followed by a Dunn's post-test for comparisons between the groups, ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Correlation between experimental variable were statistically analysed, and interpreted in regards to their Pearson correlation coefficient (ρ) and p-value.

Article Snippet: To measure anaphylatoxin C5a concentrations in the BAL and the lungs, we used a C5a mouse ELISA kit (#DY2150, R&D Systems, United States) following the manufacturer's instructions.

Techniques: In Vivo, Activation Assay, Infection, Serial Dilution, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: eBioMedicine

Article Title: Directed-complement killing of Pseudomonas aeruginosa protects against lethal pneumonia

doi: 10.1016/j.ebiom.2025.105926

Figure Lengend Snippet: CoMiX in vivo protection is associated with transient neutrophil upregulation and improved control of lung inflammation. Mice were infected/treated as described in a, sacrificed at 4 h, 8 h and 16 h p.i. , and lungs and BAL were collected for analysis. As a reflection of lung inflammation, immune cell populations were assessed by flow cytometry. The absolute number of (a) leukocytes (CD45 + cells), (b) alveolar macrophages (CD45 + SiglecF + CD11b − ), (c) eosinophils (CD45 + SiglecF + CD11c + ), and (d) neutrophils (CD45 + SiglecF − Ly6G + CD11b + ) were quantified in the BAL. (e) CD11b expression on neutrophils was measured to assess their activation. (f) Concentration of local anaphylatoxin C5a was correlated with neutrophil number in the BAL at 8 h p.i. The production of the cytokines and chemokines (g) IL-6, (h) TNF-α, and (i) CXCL-1 were quantified in the BAL by a MSD assay. All data are shown as individual values and quoted as the mean values ± SEM and the results correspond to one experiment per time-point ( n = 7–8 mice per group). Unless otherwise stated, all statistical analyses were performed using Kruskal–Wallis test followed by a Dunn's post-test for comparisons between the groups, ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. IL = interleukin; TNF = tumour necrosis factor. Correlation between experimental variable were statistically analysed, and interpreted in regards to their Pearson correlation coefficient (ρ) and p-value.

Article Snippet: To measure anaphylatoxin C5a concentrations in the BAL and the lungs, we used a C5a mouse ELISA kit (#DY2150, R&D Systems, United States) following the manufacturer's instructions.

Techniques: In Vivo, Control, Infection, Flow Cytometry, Expressing, Activation Assay, Concentration Assay