Journal: Cell Death & Disease

Article Title: A small molecule binding HMGB1 inhibits caspase-11-mediated lethality in sepsis

doi: 10.1038/s41419-021-03652-5

Figure Lengend Snippet: A Confocal microscopy of mouse peritoneal macrophages incubated for 4 h with DQ ovalbumin (10 μg/mL; red) alone or together with LPS-binding protein (10 μg/mL), or together with HMGB1(10 μg/mL) in the absence or presence of FeTPPS (1 μM) or PIX (1 μM) for 4 h, then stained with DAPI (blue). Scale bar: 10 μm. B Western blots for cathepsin D, Na + -K + -ATPase, Lamp1, Rab7 in the cytosolic fraction from vehicle-treated or LBP (5 μg/mL) or HMGB1 (5 μg/mL) with or without FeTPPS(1 μM) or PIX (1 μM)-treated mouse peritoneal macrophages. C Flow cytometry of mouse peritoneal macrophages stained with acridine orange and then treated for 4 h with LBP (10 μg/mL) or HMGB1 (10 μg/mL) with or without FeTPPS (1 μM) Numbers above bracketed lines indicate percent cells with loss of lysosomal staining with acridine orange (excitation, 488 nm; emission, 650–690 nm). D Whole-cell patch-clamp recording of HMGB1 in the absence or presence of FeTPPS (1 μM) induced inward current across the cytoplasmic membrane in proximity to the patch-clamp of HEK293 cells at acidic conditions (pH = 5.0). Graphs show the mean ± SD of technical replicates and are representative of at least three independent experiments.

Article Snippet: Recombinant mouse LPS-binding protein (LBP) (6635-LP-025/CF) was from R&D Systems Inc.

Techniques: Confocal Microscopy, Incubation, Binding Assay, Staining, Western Blot, Flow Cytometry, Patch Clamp, Membrane

Journal: Shock

Article Title: Lipopolysaccharide-Induced Cytokine Secretion from In Vitro Mouse Slow and Fast Limb Muscle

doi: 10.1097/shk.0000000000001891

Figure Lengend Snippet: FIG. 1. Representative cytokines secreted from isolated SOL in response to 1 mg/mL LPS dissolved in Krebs buffer (n ¼ 8 per group), Stats are Wilcoxon Signed ranks for matched pairs *P < 0.05, ** P < 0.01. T0 ¼ the beginning of LPS exposure, T1 ¼ 1 h after LPS exposure, T2 ¼ 2 h after LPS.

Article Snippet: Tests of SOL muscles were repeated in response to 1 mg/mL LPS in the presence of 1% C57BL/6 cytokine-free plasma (IGMS-C57-N, Innovative Research, Novi, Mich), 1 mg/mL LPS þ 1 mg/mL LPS-binding protein (LBP; 6635-LP/CF, R&D Systems, Minneapolis, Minn) or Krebs buffer alone.

Techniques: Isolation

Journal: Shock

Article Title: Lipopolysaccharide-Induced Cytokine Secretion from In Vitro Mouse Slow and Fast Limb Muscle

doi: 10.1097/shk.0000000000001891

Figure Lengend Snippet: FIG. 2. Representative cytokine secretory responses in isolated mouse SOL and mouse extensor digitorum longus (EDL) in response to 1 mg/mL LPS in Krebs buffer þ 1% serum. Stats used Wilcoxon- Signed ranks for matched pairs from time 0 to T1, and T1 to T2. ** ¼ P < 0.01, *** P < 0.001, n ¼ 10 for SOL, n ¼ 17 for EDL. T0 ¼ the beginning of LPS exposure, T1 ¼ 1 h after LPS exposure, T2 ¼ 2 h after LPS.

Article Snippet: Tests of SOL muscles were repeated in response to 1 mg/mL LPS in the presence of 1% C57BL/6 cytokine-free plasma (IGMS-C57-N, Innovative Research, Novi, Mich), 1 mg/mL LPS þ 1 mg/mL LPS-binding protein (LBP; 6635-LP/CF, R&D Systems, Minneapolis, Minn) or Krebs buffer alone.

Techniques: Isolation

Journal: Shock

Article Title: Lipopolysaccharide-Induced Cytokine Secretion from In Vitro Mouse Slow and Fast Limb Muscle

doi: 10.1097/shk.0000000000001891

Figure Lengend Snippet: FIG. 4. Comparisons of total cytokine secretion (log scale) of SOL vs. EDL at T2. Muscles were incubated in Krebs þ 1% plasma. T2 ¼ accumulated cytokines at the end of the 2 h exposure to LPS. Means SD, n ¼ 11 for SOL, n ¼ 16 for EDL. Comparisons are two sample t for parametric and Mann–Whitney for nonparametric samples. * ¼ P < 0.05, ** P < 0.01

Article Snippet: Tests of SOL muscles were repeated in response to 1 mg/mL LPS in the presence of 1% C57BL/6 cytokine-free plasma (IGMS-C57-N, Innovative Research, Novi, Mich), 1 mg/mL LPS þ 1 mg/mL LPS-binding protein (LBP; 6635-LP/CF, R&D Systems, Minneapolis, Minn) or Krebs buffer alone.

Techniques: Muscles, Incubation, Clinical Proteomics, MANN-WHITNEY

Journal: Journal of Lipid Research

Article Title: Assessment of cholesteryl ester transfer protein inhibitors for interaction with proteins involved in the immune response to infection [S]

doi: 10.1194/jlr.M002295

Figure Lengend Snippet: Effects of CETP on LPS binding to LBP and BPI. Purified CETP was added to wells containing immobilized LBP or BPI followed by addition of biotinylated LPS to achieve the final CETP concentrations indicated. Results for LPS binding to LBP (A) and BPI (B) represent the mean for four experiments each. * For effects on LPS-LBP binding at 10 nM CETP relative to control P = 0.045.

Article Snippet: Purified recombinant human LBP was purchased from R and D Systems.

Techniques: Binding Assay, Purification, Control

Journal: BMC Immunology

Article Title: Serum lipoproteins attenuate macrophage activation and Toll-Like Receptor stimulation by bacterial lipoproteins

doi: 10.1186/1471-2172-11-46

Figure Lengend Snippet: Effect of human plasma apoA-I, apoB, recombinant apoE2, apoE3, and LBP on TNF-α and IL-8 productions by primary human differentiated macrophages in response to Mip . Human monocytes/macrophages (5 × 10 5 cells/0.5 ml per well) were differentiated in a 6-day culture in the presence of 2 ng/ml M-CSF. Recombinant Mip at 1 μg/ml was exposed for 30 min to apolipoproteins or LBP before addition to macrophages. After 4 h stimulation, supernatants were collected and their content in TNF-α and IL-8 were analyzed by ELISA. Each value represents the mean ± SD of triplicates from two experiments. The results are expressed as percent of TNF-α or IL-8 release in the absence of serum. ***: p < 0.0001; **: p < 0.005, *: p < 0.05 determined by comparison with medium alone using Student's t test.

Article Snippet: Recombinant LBP was obtained from Biometec (Greifswald, Germany).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay