Journal: Critical Care

Article Title: The protective effect of apolipoprotein H in paediatric sepsis

doi: 10.1186/s13054-024-04809-2

Figure Lengend Snippet: APOH levels are significantly decreased in patients with sepsis. a LC–MS/MS was employed to identify potential biomarkers for sepsis in a pilot cohort, and ELISAs were used to confirm the expression of APOH in a validation cohort. A murine model of CLP-induced sepsis was employed to evaluate the therapeutic impact of APOH. b LC‒MS/MS was employed to identify the level of APOH in patients with sepsis and healthy controls in the pilot cohort. c LC‒MS/MS was employed to identify the level of APOH in survivors and non-survivors with sepsis in the pilot cohort. d Serum levels of APOH in 36 paediatric patients with sepsis and 69 healthy controls in the validation cohort. e The dynamics of APOH levels in the serum of paediatric patients with sepsis at 24, 48 and 72 h in the validation cohort. The data are presented as the means ± standard deviations (S.D.). “*” indicates a difference between groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Apolipoprotein H, APOH; liquid chromatography tandem mass spectrometry, LC‒MS/MS; enzyme-linked immunosorbent assay, ELISA.

Article Snippet: To neutralize APOH activity in non-severe CLP, mice in the CLP + anti-APOH (anti-APOH) group were injected with 10 μg of anti-mouse APOH neutralizing monoclonal antibody (Proteintech, Monoclonal Mouse IgG1, 66,074-1-Ig, China) dissolved in 100 μl of PBS.

Techniques: Liquid Chromatography with Mass Spectroscopy, Expressing, Biomarker Discovery, Liquid Chromatography, Mass Spectrometry, Enzyme-linked Immunosorbent Assay

Journal: Critical Care

Article Title: The protective effect of apolipoprotein H in paediatric sepsis

doi: 10.1186/s13054-024-04809-2

Figure Lengend Snippet: The expression of APOH in mice with CLP. a Serum levels of APOH in sham mice and mice with CLP at 24, 48, and 72 h after CLP (n = 6). b The relative protein levels of APOH in the lungs, livers, and kidneys (n = 5). The data are presented as the means ± standard deviations (S.D.). “*” indicates a difference between groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Apolipoprotein H, APOH; caecum ligation and puncture, CLP

Article Snippet: To neutralize APOH activity in non-severe CLP, mice in the CLP + anti-APOH (anti-APOH) group were injected with 10 μg of anti-mouse APOH neutralizing monoclonal antibody (Proteintech, Monoclonal Mouse IgG1, 66,074-1-Ig, China) dissolved in 100 μl of PBS.

Techniques: Expressing, Ligation

Journal: Critical Care

Article Title: The protective effect of apolipoprotein H in paediatric sepsis

doi: 10.1186/s13054-024-04809-2

Figure Lengend Snippet: Administration of recombinant APOH protein protects septic mice. a Ligation of the caecum at designated positions to establish a severe CLP model. The caecum was ligated (indicated by the dotted green line) at a 60% distance between the distal pole and the base of the caecum (yellow dotted line). b The procedures for rAPOH treatment in CLP-induced severe sepsis. c Survival of mice with CLP after treatment with BSA and rAPOH (n = 8). d Lung, liver, and kidney tissues were stained with H&E in the BSA and rAPOH groups. e Semiquantitative scores of tissues were calculated in the BSA and rAPOH groups (n = 7). ( f ) Serum cytokine levels in the BSA and rAPOH groups at 24 h after severe CLP (n = 7). g Cytokine levels in the PLF in the BSA and rAPOH groups at 24 h after severe CLP (n = 7). The data are presented as the means ± standard deviations (S.D.). “*” indicates a difference between groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Apolipoprotein H, APOH; recombinant murine APOH, rAPOH; bovine serum albumin, BSA; caecal ligation and puncture, CLP; peritoneal lavage fluid, PLF; haematoxylin and eosin staining, H&E

Article Snippet: To neutralize APOH activity in non-severe CLP, mice in the CLP + anti-APOH (anti-APOH) group were injected with 10 μg of anti-mouse APOH neutralizing monoclonal antibody (Proteintech, Monoclonal Mouse IgG1, 66,074-1-Ig, China) dissolved in 100 μl of PBS.

Techniques: Recombinant, Ligation, Staining

Journal: Critical Care

Article Title: The protective effect of apolipoprotein H in paediatric sepsis

doi: 10.1186/s13054-024-04809-2

Figure Lengend Snippet: Anti-APOH antibody worsens non-severe sepsis in mice. a Ligation of the caecum at designated positions to establish a non-severe CLP model. The caecum was ligated (indicated by the dotted green line) at a 40% distance between the distal pole and the base of the caecum (yellow dotted line). b The procedures for the administration of an anti-APOH antibody in mice with CLP-induced non-severe sepsis. c Survival of mice with CLP-induced non-severe sepsis after treatment with the anti-APOH antibody (n = 8). d Lung, liver, and kidney tissues were stained with H&E in the IgG control and anti-APOH groups. e Semiquantitative scores of tissues were calculated in the IgG control and anti-APOH groups (n = 7). f Serum cytokine levels in blood in the IgG control and anti-APOH groups at 24 h after non-severe CLP (n = 7). g Cytokine levels in the PLF in the IgG control and anti-APOH groups at 24 h after non-severe CLP (n = 7). The data are presented as the means ± standard deviations (S.D.). “*” indicates a difference between groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Apolipoprotein H, APOH; caecal ligation and puncture, CLP; peritoneal lavage fluid, PLF; haematoxylin and eosin staining, H&E

Article Snippet: To neutralize APOH activity in non-severe CLP, mice in the CLP + anti-APOH (anti-APOH) group were injected with 10 μg of anti-mouse APOH neutralizing monoclonal antibody (Proteintech, Monoclonal Mouse IgG1, 66,074-1-Ig, China) dissolved in 100 μl of PBS.

Techniques: Ligation, Staining, Control

Journal: Critical Care

Article Title: The protective effect of apolipoprotein H in paediatric sepsis

doi: 10.1186/s13054-024-04809-2

Figure Lengend Snippet: APOH has minimal influence on bacterial phagocytosis and killing capacities upon Pseudomonas aeruginosa infection. a CFU levels in septic mice treated with rAPOH compared with mice treated with BSA after CLP (n = 7). b The proportions of different cell types were observed in the BSA and rAPOH groups (n = 7). c Killing rates were determined in the P.a and P.a + rAPOH groups. d Immunofluorescence was employed to detect the effect of APOH on macrophage phagocytosis. e Relative fluorescence intensity of FITC-labelled P.a in the P.a. and P.a + rAPOH groups. The data are presented as the means ± standard deviations (S.D.). “*” indicates a difference between groups. * p < 0.05, ** p < 0.01. Apolipoprotein H, APOH; recombinant APOH, rAPOH; Bacterial colony-forming unit, CFU; caecal ligation and puncture, CLP; fluorescein isothiocyanate, FITC; peritoneal macrophages, PMs; peritoneal lavage fluids, PLFs; peripheral blood mononuclear cells, PBMCs; Pseudomonas aeruginosa , P.a. ; tetramethylrhodamine-5-isothiocyanate, TRIC

Article Snippet: To neutralize APOH activity in non-severe CLP, mice in the CLP + anti-APOH (anti-APOH) group were injected with 10 μg of anti-mouse APOH neutralizing monoclonal antibody (Proteintech, Monoclonal Mouse IgG1, 66,074-1-Ig, China) dissolved in 100 μl of PBS.

Techniques: Infection, Immunofluorescence, Fluorescence, Recombinant, Ligation