Journal: Molecular Therapy

Article Title: The conserved N-terminal histidine in an engineered peptide mediates sepsis treatment efficacy via dual binding to CD14 and LPS

doi: 10.1016/j.ymthe.2025.09.033

Figure Lengend Snippet: In vivo implication of histidine replacement for peptide activity (A) Effects in mouse endotoxic shock model. (Top) Schematic overview of the experimental setup. C57BL/6 mice were stimulated intraperitoneally (i.p.) with 150 μg E. coli LPS. After 30 min, 10 μg of peptide was injected i.p. into the mice. Cytokine release in plasma was quantified by flow cytometry 20 h post-LPS stimulation. (Bottom) Heatmap showing cytokine release in mice following endotoxin shock. Cytokines were detected using flow cytometry. Colors represent mean values in pg mL −1 , and each box displays mean ± SEM. (B and C) (Top) Schematic overview of the experimental setup of the polymicrobial sepsis model, followed by in vivo imaging (B) or survival (C). Polymicrobial sepsis in BALB/c tg(NF-κB-RE-Luc)-Xen reporter mice (B) or C57BL/6 (C) was induced by subjecting mice to the CLP procedure. At 30 min, the mice were i.p. treated twice daily with 100 μg peptide (in 100 μL water) for different lengths of time. The first dosage of peptide was given 30 min after the CLP procedure. The control group was given 100 μL water i.p. (B) Noninvasive in vivo bioimaging of NF-κB reporter gene expression was performed using the IVIS Spectrum system. Representative images show bioluminescence at 6, 12, and 24 h after CLP procedure. A bar chart shows measured bioluminescence intensity emitted from these reporter mice. Data are presented as means ± SEMs ( n = 5). The p values were determined using a nonparametric test followed by Dunn’s multiple comparisons test. (C) Kaplan-Meier survival curve of the percentage survival within 5 days after CLP. The p values were determined using the Kaplan-Meier method ( n = 11 mice/group). Images of mice and cell membranes were obtained from BioRender.com .

Article Snippet: BALB/c tg(NF-κB-RE-Luc)-Xen reporter mice (Taconic, 10–12 weeks old) or male C57BL/6 mice (Janvier, 8–9 weeks old) were used for the study.

Techniques: In Vivo, Activity Assay, Injection, Clinical Proteomics, Flow Cytometry, In Vivo Imaging, Control, Gene Expression