Journal: Signal Transduction and Targeted Therapy
Article Title: Acyl-CoA-binding protein (ACBP): a poor-prognosis biomarker in sepsis and a target for disease mitigation
doi: 10.1038/s41392-026-02670-z
Figure Lengend Snippet: ACBP/DBI blockade increases resistance to E. coli infection. a Twelve-week-old male C57BL/6J mice were treated with monoclonal antibody against ACBP/DBI (α-ACBP; 5 mg/kg, i.p.) twice during the week prior to E. coli challenge, followed by two additional doses (2.5 mg/kg, i.p.) at 4 h and 1 h before infection. b Survival was monitored in mice ( n = 10 per group) challenged intraperitoneally with 2.5 × 10⁶ CFU/mouse of E. coli . Statistical significance was assessed using the log-rank test. c Blood was collected at 1 h, 6 h, and 24 h post-infection (1 × 10⁶ CFU/mouse), plated on agar, and incubated overnight (O/N) at 37 °C for CFU counting ( n = 4–8 mice per group). At 6 h and 24 h post-infection, d PLF, e spleens, and f kidneys were collected and processed for CFU quantification. g Mice were pre-treated with α-ACBP as in ( a ) and additionally injected with clodronate liposomes (100 µL/10 g body weight, i.v.) or control liposomes 48 h before infection, and anti-mouse Ly6G (clone 1A8, 50 µg/mouse) or isotype control IgG2a 36 h and 12 h prior to E. coli injection. Two doses of α-ACBP (2.5 mg/kg) were also administered prior to infection ( n = 7–18 mice per group). At 24 h post-infection, h spleens and i kidneys were collected for CFU analysis. j Twelve-week-old male C57BL/6J mice ( n = 15 mice per group) were challenged with E. coli , and 30 min after treated with the monoclonal antibody against ACBP/DBI (α-ACBP; 10 mg/kg, i.p.). k At 36 h post-infection, spleens were collected and processed for CFU quantification. l Bone marrow-derived neutrophils (BMDNs) or bone marrow-derived macrophages (BMDMs) were isolated from male C57BL/6J mice and treated with α-ACBP (5 µg/mL) or IgG2a control for 16 h. m A bacterial killing assay was performed with BMDNs (≥80% Ly6G⁺), and E. coli (MOI = 10); CFU/mL were quantified the following day ( n = 5 mice per group). n Killing efficiency of BMDNs was calculated after 45 min of bacterial clearance. o BMDMs were differentiated over 7 days (≥70–80% F4/80⁺), then stimulated with LPS (100 ng/mL) + IFNγ (25 ng/mL) or IL-4 (25 ng/mL) + IL-13 (25 ng/mL) for 24 h, and treated with α-ACBP or IgG2a control for 16 h ( n = 6 mice per group). Killing assays were performed with E. coli (MOI = 10), and CFU/mL were determined after overnight incubation. p Bacterial killing by BMDMs was quantified after 120 min and expressed as absolute values of [CFU/mL/min]. q BMDMs from male C57BL/6J and Gabrg2 F77I/F77I mice were differentiated over 7 days (≥70–80% F4/80⁺), then stimulated with LPS (100 ng/mL) + IFNγ (25 ng/mL) or IL-4 (25 ng/mL) + IL-13 (25 ng/mL) for 24 h ( n = 6 mice per group). Killing assays were performed with E. coli (MOI = 10), and bacterial killing by BMDMs was quantified after 120 min and expressed as absolute values of CFU/mL/min. Data are presented as means ± SEM. Statistical comparisons were performed using one-way or two-way ANOVA with estimation of marginal means for pairwise comparisons, or Student’s t-test where applicable. l was generated with “BioRender.com.” IFNγ interferon-γ, IL-4 interleukin-4, IL-13 interleukin-13, MOI multiplicity of infection, CFU colony-forming units, PLF peritoneal lavage fluid
Article Snippet: Neutrophil depletion was achieved by intraperitoneal injections of anti-mouse Ly6G antibody (clone 1A8, 50 μg/mouse, #BE0075-1, BioXcell) or isotype IgG2a control (clone 2A3, #BE0089, BioXcell) at 36 h and 12 h before E. coli injection., Additionally, α-ACBP (2.5 mg/kg) was administered twice more in the hours preceding E. coli challenge.
Techniques: Infection, Incubation, Injection, Liposomes, Control, Derivative Assay, Isolation, Generated
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![ACBP/DBI blockade increases resistance to E. coli infection. a Twelve-week-old male C57BL/6J mice were treated with monoclonal antibody against ACBP/DBI (α-ACBP; 5 mg/kg, i.p.) twice during the week prior to E. coli challenge, followed by two additional doses (2.5 mg/kg, i.p.) at 4 h and 1 h before infection. b Survival was monitored in mice ( n = 10 per group) challenged intraperitoneally with 2.5 × 10⁶ CFU/mouse of E. coli . Statistical significance was assessed using the log-rank test. c Blood was collected at 1 h, 6 h, and 24 h post-infection (1 × 10⁶ CFU/mouse), plated on agar, and incubated overnight (O/N) at 37 °C for CFU counting ( n = 4–8 mice per group). At 6 h and 24 h post-infection, d PLF, e spleens, and f kidneys were collected and processed for CFU quantification. g Mice were pre-treated with α-ACBP as in ( a ) and additionally injected with clodronate liposomes (100 µL/10 g body weight, i.v.) or control liposomes 48 h before infection, and anti-mouse Ly6G (clone 1A8, 50 µg/mouse) or isotype control IgG2a 36 h and 12 h prior to E. coli injection. Two doses of α-ACBP (2.5 mg/kg) were also administered prior to infection ( n = 7–18 mice per group). At 24 h post-infection, h spleens and i kidneys were collected for CFU analysis. j Twelve-week-old male C57BL/6J mice ( n = 15 mice per group) were challenged with E. coli , and 30 min after treated with the monoclonal antibody against ACBP/DBI (α-ACBP; 10 mg/kg, i.p.). k At 36 h post-infection, spleens were collected and processed for CFU quantification. l Bone marrow-derived neutrophils (BMDNs) or bone marrow-derived macrophages (BMDMs) were isolated from male C57BL/6J mice and treated with α-ACBP (5 µg/mL) or IgG2a control for 16 h. m A bacterial killing assay was performed with BMDNs (≥80% Ly6G⁺), and E. coli (MOI = 10); CFU/mL were quantified the following day ( n = 5 mice per group). n Killing efficiency of BMDNs was calculated after 45 min of bacterial clearance. o BMDMs were differentiated over 7 days (≥70–80% F4/80⁺), then stimulated with LPS (100 ng/mL) + IFNγ (25 ng/mL) or IL-4 (25 ng/mL) + IL-13 (25 ng/mL) for 24 h, and treated with α-ACBP or IgG2a control for 16 h ( n = 6 mice per group). Killing assays were performed with E. coli (MOI = 10), and CFU/mL were determined after overnight incubation. p Bacterial killing by BMDMs was quantified after 120 min and expressed as absolute values of [CFU/mL/min]. q BMDMs from male C57BL/6J and Gabrg2 F77I/F77I mice were differentiated over 7 days (≥70–80% F4/80⁺), then stimulated with LPS (100 ng/mL) + IFNγ (25 ng/mL) or IL-4 (25 ng/mL) + IL-13 (25 ng/mL) for 24 h ( n = 6 mice per group). Killing assays were performed with E. coli (MOI = 10), and bacterial killing by BMDMs was quantified after 120 min and expressed as absolute values of CFU/mL/min. Data are presented as means ± SEM. Statistical comparisons were performed using one-way or two-way ANOVA with estimation of marginal means for pairwise comparisons, or Student’s t-test where applicable. l was generated with “BioRender.com.” IFNγ interferon-γ, IL-4 interleukin-4, IL-13 interleukin-13, MOI multiplicity of infection, CFU colony-forming units, PLF peritoneal lavage fluid](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6771/pmc13046771/pmc13046771__41392_2026_2670_Fig3_HTML.jpg)
