Journal:

Article Title: Endothelial Adhesion Molecule Expression and Its Inhibition by Recombinant Bactericidal/Permeability-Increasing Protein Are Influenced by the Capsulation and Lipooligosaccharide Structure of Neisseria meningitidis

doi:

Figure Lengend Snippet: Differential induction of CD62E on HUVEC by the N. meningitidis B1940 parent and the cpsD and siaD mutants occurs over a range of bacterial concentrations. Bacteria were added at 108 to 104 organisms per ml. After 15 min of incubation, HUVEC were washed thoroughly in fresh medium and incubated for a further 5 h. CD62E expression was determined at 5 h by flow cytometry. This is representative of at least three experiments yielding similar results.

Article Snippet: Adhesion molecule expression was detected by incubation with mouse monoclonal antibodies to human ICAM-1, CD62E, and VCAM-1 (Serotec, Oxford, United Kingdom) followed by goat anti-mouse F(ab′) 2 -phycoerythrin conjugate (Dako, High Wycombe, United Kingdom).

Techniques: Incubation, Expressing, Flow Cytometry

Journal:

Article Title: Endothelial Adhesion Molecule Expression and Its Inhibition by Recombinant Bactericidal/Permeability-Increasing Protein Are Influenced by the Capsulation and Lipooligosaccharide Structure of Neisseria meningitidis

doi:

Figure Lengend Snippet: Flow cytometric analysis of expression of CD62E, ICAM-1, and VCAM-1 on HUVEC following 5 h of incubation with 100 ng of E. coli O111:B4 LPS, 100 ng of meningococcal LOS, and 107 N. meningitidis B1940 parent and siaD mutant organisms per ml. Shaded areas represent CD62E, ICAM-1, and VCAM-1 staining in response to stimuli; the solid line represents cell adhesion molecule staining in unstimulated cells; and the dotted line represents staining with an irrelevant mouse immunoglobulin G1 antibody. Numbers on each plot are median fluorescent intensities and percent positive events for stimulated cells. Data presented here is representative of three experiments yielding similar results.

Article Snippet: Adhesion molecule expression was detected by incubation with mouse monoclonal antibodies to human ICAM-1, CD62E, and VCAM-1 (Serotec, Oxford, United Kingdom) followed by goat anti-mouse F(ab′) 2 -phycoerythrin conjugate (Dako, High Wycombe, United Kingdom).

Techniques: Expressing, Incubation, Mutagenesis, Staining

Journal:

Article Title: Endothelial Adhesion Molecule Expression and Its Inhibition by Recombinant Bactericidal/Permeability-Increasing Protein Are Influenced by the Capsulation and Lipooligosaccharide Structure of Neisseria meningitidis

doi:

Figure Lengend Snippet: Brief exposure of the unencapsulated siaD mutant to HUVEC is a potent inducer of expression of CD62E. (A) N. meningitidis B1940 siaD mutant at 106 organisms per ml or 10 ng of E. coli LPS was added to HUVEC, which were then washed thoroughly with fresh medium at 5, 15, 60, 120, and 240 min. (B) N. meningitidis B1940 siaD and parent organisms were incubated at 106 organisms per ml and washed at 15, 60, 120, and 240 min. CD62E expression was determined by flow cytometry after 5 h of incubation. The results shown here are representative experiments from at least three separate experiments that yielded similar results.

Article Snippet: Adhesion molecule expression was detected by incubation with mouse monoclonal antibodies to human ICAM-1, CD62E, and VCAM-1 (Serotec, Oxford, United Kingdom) followed by goat anti-mouse F(ab′) 2 -phycoerythrin conjugate (Dako, High Wycombe, United Kingdom).

Techniques: Mutagenesis, Expressing, Incubation, Flow Cytometry

Journal:

Article Title: Endothelial Adhesion Molecule Expression and Its Inhibition by Recombinant Bactericidal/Permeability-Increasing Protein Are Influenced by the Capsulation and Lipooligosaccharide Structure of Neisseria meningitidis

doi:

Figure Lengend Snippet: Effect of rBPI21 on E. coli O111:B4 LPS-induced HUVEC expression of CD62E, ICAM-1, and VCAM-1. E. coli LPS (100 ng/ml) was added to HUVEC preincubated with 10 μg of rBPI21 per ml. Cell adhesion molecule determination was determined by flow cytometry after 5 h of incubation. The results shown are representative of multiple experiments with same level of inhibition.

Article Snippet: Adhesion molecule expression was detected by incubation with mouse monoclonal antibodies to human ICAM-1, CD62E, and VCAM-1 (Serotec, Oxford, United Kingdom) followed by goat anti-mouse F(ab′) 2 -phycoerythrin conjugate (Dako, High Wycombe, United Kingdom).

Techniques: Expressing, Flow Cytometry, Incubation, Inhibition

Journal:

Article Title: Endothelial Adhesion Molecule Expression and Its Inhibition by Recombinant Bactericidal/Permeability-Increasing Protein Are Influenced by the Capsulation and Lipooligosaccharide Structure of Neisseria meningitidis

doi:

Figure Lengend Snippet: Effect of rBPI21 on HUVEC expression of CD62E by various doses of the N. meningitidis B1940 siaD mutant. rBPI21 (10 μg/ml) or medium was added to HUVEC, and then either 100 ng of E. coli O111:B4 LPS or various concentrations of bacteria were added as shown. CD62E expression was determined after 5 h. The data presented here are histograms of fluorescence intensity of phycoerythrin fluorochrome plotted against the number of events. The dotted line represents resting CD62E expression; the shaded area represents CD62E expression with LPS or bacteria; and the continuous black line represents CD62E expression when LPS or bacteria were preincubated with rBPI21.

Article Snippet: Adhesion molecule expression was detected by incubation with mouse monoclonal antibodies to human ICAM-1, CD62E, and VCAM-1 (Serotec, Oxford, United Kingdom) followed by goat anti-mouse F(ab′) 2 -phycoerythrin conjugate (Dako, High Wycombe, United Kingdom).

Techniques: Expressing, Mutagenesis, Fluorescence

Journal:

Article Title: Endothelial Adhesion Molecule Expression and Its Inhibition by Recombinant Bactericidal/Permeability-Increasing Protein Are Influenced by the Capsulation and Lipooligosaccharide Structure of Neisseria meningitidis

doi:

Figure Lengend Snippet: Effect of rBPI21 on induction of CD62E expression on HUVEC in response to N. meningitidis B1940 parent and cps, siaD, and cpsD mutants (A) and B1940 parent and the lst mutant (7B). HUVEC were pre-incubated with 10 μg of rBPI21 per ml, and then 105 organisms/ml were added. CD62E expression was measured after 5 h by flow cytometry. The results shown are the mean median fluorescence intensity (MFI) and standard error of the mean from three separate experiments.

Article Snippet: Adhesion molecule expression was detected by incubation with mouse monoclonal antibodies to human ICAM-1, CD62E, and VCAM-1 (Serotec, Oxford, United Kingdom) followed by goat anti-mouse F(ab′) 2 -phycoerythrin conjugate (Dako, High Wycombe, United Kingdom).

Techniques: Expressing, Mutagenesis, Incubation, Flow Cytometry, Fluorescence

Journal:

Article Title: Endothelial Adhesion Molecule Expression and Its Inhibition by Recombinant Bactericidal/Permeability-Increasing Protein Are Influenced by the Capsulation and Lipooligosaccharide Structure of Neisseria meningitidis

doi:

Figure Lengend Snippet: Effect of time exposure of rBPI21 on induction of CD62E on HUVEC after 5 h of stimulation with 10 ng of E. coli O111:B4 LPS or N. meningitidis B1940 parent and siaD mutant strains. Bacteria were added at 106 per ml and washed off after 15 min. Then 5 μg of rBPI21 was added at 15, 60, and 120 min. E. coli O111:B4 LPS (10 ng) was added, and 5 μg of rBPI21 was added at the same time points. The results shown are a representative of three experiments with similar results.

Article Snippet: Adhesion molecule expression was detected by incubation with mouse monoclonal antibodies to human ICAM-1, CD62E, and VCAM-1 (Serotec, Oxford, United Kingdom) followed by goat anti-mouse F(ab′) 2 -phycoerythrin conjugate (Dako, High Wycombe, United Kingdom).

Techniques: Mutagenesis