human c1-esterase inhibitor  (CSL Behring)

Journal: Scientific Reports

Article Title: Evidence, detailed characterization and clinical context of complement activation in acute multisystem inflammatory syndrome in children

doi: 10.1038/s41598-022-23806-5

Figure Lengend Snippet: Key resources.

Article Snippet: Polyclonal antiserum to human C1 inhibitor (C1 esterase) , Quidel , Cat# A300.

Techniques: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Software

Journal: Immunologic Research

Article Title: Interplay between C1-inhibitor and group IIA secreted phospholipase A 2 impairs their respective function

doi: 10.1007/s12026-022-09331-7

Figure Lengend Snippet: Effects of rhC1-INH and pdC1-INH on secretion of IL-6, TNF-α, and CXCL8 from human PBMCs. PBMCs were stimulated (16 h, 37 °C) with or without the indicated concentrations of rhC1-INH or pdC1-INH. IL-6 ( a ), TNF-α ( b ), and CXCL8 ( c ) concentrations in cell medium were determined by ELISA and values were normalized relative to the amount of total proteins in each well (measured by Bradford assay on cell lysates). The data are reported as mean ± SEM of six different preparations of PBMCs from six different donors

Article Snippet: The following reagents were purchased: l -glutamine, antibiotic–antimycotic solution (10,000 IU/mL penicillin, 10 mg/mL streptomycin, and 25 µg/mL amphotericin B), detoxified LPS (from E. coli serotype 0111:B4), polymyxin B sulfate (Sigma-Aldrich, Milan, Italy), RPMI and fetal calf serum (FCS, endotoxin level < 0.1EU/mL, MP Biomedicals Europe, Illkirch, France), recombinant human C1-esterase inhibitor (rhC1-INH, PeproTech, USA), human plasma–derived C1-esterase inhibitor (pdC1-INH; Cinryze®, Takeda Pharmaceutical Company, Tokyo, Japan).

Techniques: Enzyme-linked Immunosorbent Assay, Bradford Assay

Journal: Immunologic Research

Article Title: Interplay between C1-inhibitor and group IIA secreted phospholipase A 2 impairs their respective function

doi: 10.1007/s12026-022-09331-7

Figure Lengend Snippet: Effect of rhC1-INH and pdC1-INH on hGIIA- or LPS-induced release of IL-6, TNF-α, and CXCL8 from human PBMCs. PBMCs were stimulated (16 h, 37 °C) with hGIIA ( a – c ) or LPS ( d – f ) alone (white column) or in combination with rhC1-INH (light gray) or pdC1-INH (dark gray). IL-6 ( a , d ), TNF-α ( b , e ), and CXCL8 ( c , f ) release was determined by ELISA and values were normalized relative to the amount of total proteins in each well (measured by Bradford assay on cell lysates). The data are reported as mean ± SEM of six different preparations of PBMCs from six different donors. * p < 0.05 vs. ** p < 0.01 and *** p < 0.001 vs. control (black column). § p < 0.05 vs. §§ p < 0.01 and. §§§ p < 0.001 vs. hGIIA, or LPS alone (white column)

Article Snippet: The following reagents were purchased: l -glutamine, antibiotic–antimycotic solution (10,000 IU/mL penicillin, 10 mg/mL streptomycin, and 25 µg/mL amphotericin B), detoxified LPS (from E. coli serotype 0111:B4), polymyxin B sulfate (Sigma-Aldrich, Milan, Italy), RPMI and fetal calf serum (FCS, endotoxin level < 0.1EU/mL, MP Biomedicals Europe, Illkirch, France), recombinant human C1-esterase inhibitor (rhC1-INH, PeproTech, USA), human plasma–derived C1-esterase inhibitor (pdC1-INH; Cinryze®, Takeda Pharmaceutical Company, Tokyo, Japan).

Techniques: Enzyme-linked Immunosorbent Assay, Bradford Assay

Journal: Immunologic Research

Article Title: Interplay between C1-inhibitor and group IIA secreted phospholipase A 2 impairs their respective function

doi: 10.1007/s12026-022-09331-7

Figure Lengend Snippet: Effects of hGIIA on C1-INH activity. a Plasma from normal donors was preincubated (2 h, 37 °C) with or without hGIIA (3 µg/mL) or LPS (100 ng/mL) and the functional activity of C1-INH was then evaluated by a colorimetric assay. Data are expressed as percent inhibition of the maximum plasma activity of C1-INH calculated as ( R − R b ) × 100, where R is the C1-INH activity in plasma samples treated with the hGIIA or LPS, and R b is the C1-INH activity in unstimulated samples. ** p < 0.01 vs. control. b rhC1-INH and pdC1-INH were incubated (2 h, 37 °C) with or without hGIIA and then functional activity of C1-esterase was evaluated by colorimetric assay. ** p < 0.01 vs. rhC1-INH or pdC1-INH alone. c hGIIA was preincubated with RO032107A and then incubated with or without rhC1-INH or pdC1-INH (2 h, 37 °C) after which the functional activity of C1-esterase was evaluated. Data are the mean ± SD of 3 experiments. ** p < 0.01 vs. rhC1-INH or pdC1-INH alone

Article Snippet: The following reagents were purchased: l -glutamine, antibiotic–antimycotic solution (10,000 IU/mL penicillin, 10 mg/mL streptomycin, and 25 µg/mL amphotericin B), detoxified LPS (from E. coli serotype 0111:B4), polymyxin B sulfate (Sigma-Aldrich, Milan, Italy), RPMI and fetal calf serum (FCS, endotoxin level < 0.1EU/mL, MP Biomedicals Europe, Illkirch, France), recombinant human C1-esterase inhibitor (rhC1-INH, PeproTech, USA), human plasma–derived C1-esterase inhibitor (pdC1-INH; Cinryze®, Takeda Pharmaceutical Company, Tokyo, Japan).

Techniques: Activity Assay, Functional Assay, Colorimetric Assay, Inhibition, Incubation

Journal: Immunologic Research

Article Title: Interplay between C1-inhibitor and group IIA secreted phospholipase A 2 impairs their respective function

doi: 10.1007/s12026-022-09331-7

Figure Lengend Snippet: Effect of rhC1-INH and pdC1-INH on hGIIA enzymatic activity. hGIIA (10 pM) was preincubated with the indicated concentrations of recombinant and plasma-derived C1-INH in 100 µL of sPLA 2 activity buffer for 15 min at room temperature. sPLA 2 enzymatic activity was measured as reported in the “ ” section

Article Snippet: The following reagents were purchased: l -glutamine, antibiotic–antimycotic solution (10,000 IU/mL penicillin, 10 mg/mL streptomycin, and 25 µg/mL amphotericin B), detoxified LPS (from E. coli serotype 0111:B4), polymyxin B sulfate (Sigma-Aldrich, Milan, Italy), RPMI and fetal calf serum (FCS, endotoxin level < 0.1EU/mL, MP Biomedicals Europe, Illkirch, France), recombinant human C1-esterase inhibitor (rhC1-INH, PeproTech, USA), human plasma–derived C1-esterase inhibitor (pdC1-INH; Cinryze®, Takeda Pharmaceutical Company, Tokyo, Japan).

Techniques: Activity Assay, Recombinant, Derivative Assay

Journal: Immunologic Research

Article Title: Interplay between C1-inhibitor and group IIA secreted phospholipase A 2 impairs their respective function

doi: 10.1007/s12026-022-09331-7

Figure Lengend Snippet: Binding assays for C1-INH-hGIIA interaction: SPR and intrinsic fluorescence. Overlay of a sensorgrams recorded at increasing concentrations of C1-INH on hGIIA-chip. b Fluorescence emission spectra of C1-INH at increasing equivalents of hGIIA

Article Snippet: The following reagents were purchased: l -glutamine, antibiotic–antimycotic solution (10,000 IU/mL penicillin, 10 mg/mL streptomycin, and 25 µg/mL amphotericin B), detoxified LPS (from E. coli serotype 0111:B4), polymyxin B sulfate (Sigma-Aldrich, Milan, Italy), RPMI and fetal calf serum (FCS, endotoxin level < 0.1EU/mL, MP Biomedicals Europe, Illkirch, France), recombinant human C1-esterase inhibitor (rhC1-INH, PeproTech, USA), human plasma–derived C1-esterase inhibitor (pdC1-INH; Cinryze®, Takeda Pharmaceutical Company, Tokyo, Japan).

Techniques: Binding Assay, Fluorescence

Journal: Immunologic Research

Article Title: Interplay between C1-inhibitor and group IIA secreted phospholipase A 2 impairs their respective function

doi: 10.1007/s12026-022-09331-7

Figure Lengend Snippet: C1-INH:hGIIA complex. A Docking poses: predicted conformations of hGIIA (gray shades) on C1-INH (green); the lowest scoring hGIIA conformation is highlighted (cyan). Interacting residues are indicated (licorice) with aromatic side chains further highlighted by their van der Waals spheres. Aromatic residues not involved in the interaction are also indicated (yellow). B Schematic diagram of the interaction between C1-INH (top, green shade) and hGIIA (bottom, blue shade) in the optimum HADDOCK pose of panel A . C Molecular dynamics simulation analysis: comparison between initial ( t = 0, gray shade) and final ( t = 500 ns, color) configurations of C1-INH:hGIIA. Simulations were run at T = 330 K in full water solvent. Water molecules are not shown for ease of visualization. Interacting aromatic residues are highlighted with their van der Waals spheres. Gln463, forming a hydrogen bond with Tyr66, is also explicitly drawn. In the inset: the backbone RMSD of hGIIA (blue), of C1-INH (green), and of hGIIA with respect to C1-INH (black). D Schematic diagram of the interaction between C1-INH (top, green shade) and hGIIA (bottom, blue shade) in final ( t = 500 ns) configurations of panel C . In A and D , all residues interacting with the target are listed; hydrogen bonds are indicated by dotted lines (magenta). The hydrogen bond distances are also indicated. Aromatic residues are highlighted (dashed circles)

Article Snippet: The following reagents were purchased: l -glutamine, antibiotic–antimycotic solution (10,000 IU/mL penicillin, 10 mg/mL streptomycin, and 25 µg/mL amphotericin B), detoxified LPS (from E. coli serotype 0111:B4), polymyxin B sulfate (Sigma-Aldrich, Milan, Italy), RPMI and fetal calf serum (FCS, endotoxin level < 0.1EU/mL, MP Biomedicals Europe, Illkirch, France), recombinant human C1-esterase inhibitor (rhC1-INH, PeproTech, USA), human plasma–derived C1-esterase inhibitor (pdC1-INH; Cinryze®, Takeda Pharmaceutical Company, Tokyo, Japan).

Techniques: