Journal: Thorax

Article Title: Interferon gamma applied ex vivo restores function to neutrophils from critically ill patients

doi: 10.1136/thorax-2025-223280

Figure Lengend Snippet: Neutrophil phagocytosis in critically ill patients is improved by IFN-γ treatment. ( A, B ) Neutrophil phagocytosis was measured and patients characterised on the basis of whether neutrophils were functional or dysfunctional as described in the Methods section. Dysfunction is defined as <50% of neutrophils ingesting ≥2 zymosan particles. Neutrophils from patients were incubated with IFN-γ (1 ng/mL) or control for 1 hour and phagocytosis of zymosan assessed. ( C ) Neutrophils from patients with dysfunctional neutrophils were preincubated with a PI3K-γ inhibitor AS (AS 605420, 10 nM) or FcX (Fc block) for 30 min before the application of IFN-γ (1 ng/mL) for 1 hour. Bars represent medians. ( A ) Wilcoxon signed rank test (n=8). ( B ) Wilcoxon t-test (n=1). ( C ) Friedman test with Dunn’s multiple comparisons test (n=6). Comparisons shown are against the second (IFN-γ) column.

Article Snippet: Recombinant human IFN-γ was purchased from R&D Systems (Abingdon, UK).

Techniques: Functional Assay, Incubation, Control, Blocking Assay

Journal: Thorax

Article Title: Interferon gamma applied ex vivo restores function to neutrophils from critically ill patients

doi: 10.1136/thorax-2025-223280

Figure Lengend Snippet: IFN-γ improves bacterial killing in dysfunctional neutrophils from critically ill patients. Percentage of Staphylococcus aureus killed by neutrophils isolated from seven critically ill patients with neutrophil dysfunction. Dysfunctional neutrophils were preincubated with Fc block (FcX) or a PI3K-γ inhibitor (AS605420 or ‘AS’, 10 nM) for 30 min before the application of IFN-γ (1 ng/mL) for 1 hour. Friedman test with Dunn’s multiple comparisons test. Bars represent medians. Comparisons shown are against the second (IFN-γ) column.

Article Snippet: Recombinant human IFN-γ was purchased from R&D Systems (Abingdon, UK).

Techniques: Isolation, Blocking Assay

Journal: Thorax

Article Title: Interferon gamma applied ex vivo restores function to neutrophils from critically ill patients

doi: 10.1136/thorax-2025-223280

Figure Lengend Snippet: IFN-γ increases intracellular ROS release in dysfunctional neutrophils from critically ill patients. Whole blood was incubated with 1 µM DHR123 before incubation with 10 nM AS605420 for 30 min prior to addition of 1 ng/mL IFN-γ for 1 hour and subsequent stimulation with 1 µM PMA. ( A ) Data are expressed as fluorescence intensity and shown as individual points from 7 patients. Bars represent medians. Friedman test with Dunn’s multiple comparisons test. ( B ) Representative histogram of DHR fluorescence generated using FCS Express. DHR, dihydrorhodamine; MedFL, median fluorescence intensity; PMA, phorbol myristate acetate; ROS, reactive oxygen species.

Article Snippet: Recombinant human IFN-γ was purchased from R&D Systems (Abingdon, UK).

Techniques: Incubation, Fluorescence, Generated

Journal: Thorax

Article Title: Interferon gamma applied ex vivo restores function to neutrophils from critically ill patients

doi: 10.1136/thorax-2025-223280

Figure Lengend Snippet: Cell surface expression of phagocytic receptors on dysfunctional neutrophils after incubation of whole blood with IFN-γ. Expression of neutrophil CD64 ( A ), CD16 ( B ), CD32 ( C ) and CD11b ( D ) was measured by flow cytometry following incubation of whole blood with IFN-γ (1 ng/mL) for 1 hour. Wilcoxon signed rank test, n=7. ( E ) Representative fluorescence histogram for each marker, generated using FCS Express. MedFL, median fluorescence intensity.

Article Snippet: Recombinant human IFN-γ was purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing, Incubation, Flow Cytometry, Fluorescence, Marker, Generated

Journal: Thorax

Article Title: Interferon gamma applied ex vivo restores function to neutrophils from critically ill patients

doi: 10.1136/thorax-2025-223280

Figure Lengend Snippet: IFN-γ increases Rac and Cdc42 activity in dysfunctional neutrophils from critically ill patients. IFN-γ (1 ng/mL for 1 hour) or vehicle was added to dysfunctional neutrophils from critically ill patients (n=6). ( A ) Cdc42 and ( C ) Rac 1,2,3 activity was then assessed by G-LISA. Results are expressed as absorbance values at 490 nm. ( B ) Total Cdc42 and ( D ) total Rac were assessed by ELISA. Wilcoxon signed rank test.

Article Snippet: Recombinant human IFN-γ was purchased from R&D Systems (Abingdon, UK).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay