Journal: Rheumatology (Oxford, England)

Article Title: Lactoferrin is a survival factor for neutrophils in rheumatoid synovial fluid

doi: 10.1093/rheumatology/ken412

Figure Lengend Snippet: Lactoferrin levels in RA SF. Lactoferrin concentrations were measured by ELISA in SFs from patients with RA of >2 yrs ( n = 26) (•) or <3 months (♦) duration ( n = 10) and in matched sera from 11 of the patients with established RA (▪). The median value is indicated by a bar.

Article Snippet: For immunodepletion of lactoferrin, SF was incubated with an anti-lactoferrin monoclonal antibody (Sigma-Aldrich) or an isotype matched control and the antibody removed using protein G linked to magnetic beads (µ beads, Miltenyi Biotec, Surrey, UK).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Rheumatology (Oxford, England)

Article Title: Lactoferrin is a survival factor for neutrophils in rheumatoid synovial fluid

doi: 10.1093/rheumatology/ken412

Figure Lengend Snippet: Effect of lactoferrin on neutrophil apoptosis and function. ( A ) Peripheral blood neutrophils were incubated with medium alone or with medium containing iron-free lactoferrin at the concentrations shown and apoptosis measured by loss of DiOC6 staining after 9 h in culture. All incubations also contained 10 μg/ml polymyxin B. Data are mean ± s.d . ( n = 5) and * P < 0.04, ** P < 0.02. ( B ) Neutrophils were incubated with 100 μg/ml lactoferrin (LTF) for 20 h prior to stimulation with fMLP and measurement of superoxide generation. Data are expressed as mean ± s.d . of the area under the curve (AUC) per 10 6 viable cells ( n = 3). * P < 0.01. ( C ) Neutrophils isolated from RA synovial fluid were incubated with medium alone, 50 ng/ml GM-CSF or 100 μg/ml lactoferrin. Apoptosis was measured by active caspase 3 staining after 9 h in culture. Data are mean ± s.d . ( n = 3) and * P < 0.05. All data were analysed by Student's t- test.

Article Snippet: For immunodepletion of lactoferrin, SF was incubated with an anti-lactoferrin monoclonal antibody (Sigma-Aldrich) or an isotype matched control and the antibody removed using protein G linked to magnetic beads (µ beads, Miltenyi Biotec, Surrey, UK).

Techniques: Incubation, Staining, Isolation

Journal: Rheumatology (Oxford, England)

Article Title: Lactoferrin is a survival factor for neutrophils in rheumatoid synovial fluid

doi: 10.1093/rheumatology/ken412

Figure Lengend Snippet: Comparison of lactoferrin effects with known neutrophil survival factors. Peripheral blood neutrophils were incubated with medium alone (control), 100 ng/ml LPS, 800 IU/ml IFN-β, 50 ng/ml GM-CSF, 100 μg/ml lactoferrin (LTF) and apoptosis measured by differential staining and nuclear morphology after 9 h in culture. Data are mean ± s.d . ( n = 5) and were analysed by ANOVA followed by Dunnett's comparison with control. * P < 0.05, ** P < 0.01.

Article Snippet: For immunodepletion of lactoferrin, SF was incubated with an anti-lactoferrin monoclonal antibody (Sigma-Aldrich) or an isotype matched control and the antibody removed using protein G linked to magnetic beads (µ beads, Miltenyi Biotec, Surrey, UK).

Techniques: Incubation, Staining

Journal: Rheumatology (Oxford, England)

Article Title: Lactoferrin is a survival factor for neutrophils in rheumatoid synovial fluid

doi: 10.1093/rheumatology/ken412

Figure Lengend Snippet: The anti-apoptotic effect of synovial fluid is reduced by depletion of lactoferrin. Peripheral blood neutrophils were incubated with medium alone (control), or with synovial fluid from seven different RA patients which had been immunodepleted for lactoferrin (LTF) using an anti-lactoferrin monoclonal antibody (LTF) or mock-depleted using an isotype control immunoglobulin (Mock). Apoptosis was measured by active caspase 3 staining after 20 h in culture. Control and immunodepleted data were compared using a paired Student's t -test and * P < 0.03.

Article Snippet: For immunodepletion of lactoferrin, SF was incubated with an anti-lactoferrin monoclonal antibody (Sigma-Aldrich) or an isotype matched control and the antibody removed using protein G linked to magnetic beads (µ beads, Miltenyi Biotec, Surrey, UK).

Techniques: Incubation, Staining

Journal: Rheumatology (Oxford, England)

Article Title: Lactoferrin is a survival factor for neutrophils in rheumatoid synovial fluid

doi: 10.1093/rheumatology/ken412

Figure Lengend Snippet: Lactoferrin levels correlate with serum CRP and RF status. Lactoferrin levels were measured in synovial fluid of 24 patients with RA and correlated with ( A ) age, ( B ) disease duration, ( C ) serum ESR, ( D ) presence of joint erosion, ( E ) serum CRP and ( F ) seropositivity for RF. Data were analysed by Spearman's correlation.

Article Snippet: For immunodepletion of lactoferrin, SF was incubated with an anti-lactoferrin monoclonal antibody (Sigma-Aldrich) or an isotype matched control and the antibody removed using protein G linked to magnetic beads (µ beads, Miltenyi Biotec, Surrey, UK).

Techniques: