Journal: British Journal of Pharmacology

Article Title: Repurposing drugs targeting the P2X7 receptor to limit hyperinflammation and disease during influenza virus infection

doi: 10.1111/bph.14787

Figure Lengend Snippet: The P2X7 receptor antagonist AZ11645373 inhibits NLRP3‐dependent release of IL‐1β in macrophages. iBMDMs were primed with LPS (100 ng·ml −1 ) for 3 hr then treated with a range of doses of AZ11645373 for 1 hr. Cells were then stimulated with NLRP3 activators PR8 PB1‐F2 peptide, silica, ATP, or nigericin at the indicated concentrations in triplicate for 6 hr. Cellular supernatants were analysed for secreted IL‐1β by ELISA. Results are representative of n = 9 and mean ± SEM. * P < .05, significantly different from NLRP3 activator alone; one‐way ANOVA

Article Snippet: Following infection, mice were treated at the time points indicated (every 48 hr; according to ethically approved guidelines) with 40 mg·kg −1 probenecid (water soluble; ThermoFisher) or 20 mg·kg −1 AZ11645373 (Torcis) via the intranasal route in 50‐μl PBS.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: British Journal of Pharmacology

Article Title: Repurposing drugs targeting the P2X7 receptor to limit hyperinflammation and disease during influenza virus infection

doi: 10.1111/bph.14787

Figure Lengend Snippet: Late administration of probenecid or AZ11645373 improves survival and disease during severe HKx31 IAV infection. Groups of C57BL/6 mice were infected intranasally with a high dose ( n = 8 per group) of HKx31 (10 5 PFU). Mice were treated intranasally with probenecid (40 mg·kg −1 ), AZ11645373 (20 mg·kg −1 ), or PBS on day 3 post‐infection and every 48 hr thereafter (arrows). Uninfected mice treated with probenecid or AZ11645373 are included for comparison. (a) Mice were weighed daily, and results were expressed as mean per cent weight change ± SEM. (b) Survival curves are shown. * P < .05 PBS, significantly different from AZ11645373; # P < .05, PBS significantly different from probenecid; Mantel–Cox log‐rank test

Article Snippet: Following infection, mice were treated at the time points indicated (every 48 hr; according to ethically approved guidelines) with 40 mg·kg −1 probenecid (water soluble; ThermoFisher) or 20 mg·kg −1 AZ11645373 (Torcis) via the intranasal route in 50‐μl PBS.

Techniques: Infection, Comparison

Journal: British Journal of Pharmacology

Article Title: Repurposing drugs targeting the P2X7 receptor to limit hyperinflammation and disease during influenza virus infection

doi: 10.1111/bph.14787

Figure Lengend Snippet: Late administration of probenecid or AZ11645373 improves survival and disease following infection with mouse‐adapted PR8 H1N1. Groups of C57BL/6 mice were infected intranasally with a lethal dose ( n = 8 per group) of PR8 (50 PFU). Mice were treated intranasally with probenecid (40 mg·kg −1 ), AZ11645373 (20 mg·kg −1 ), or PBS on day 7 post‐infection and every 48 hr thereafter (arrows). (a) Mice were weighed daily, and results are expressed as mean per cent weight change ± SEM. (b) Survival curves are shown. * P < .05, PBS significantly different from AZ11645373; # P < .05, PBS significantly different from probenecid, Mantel–Cox log‐rank test

Article Snippet: Following infection, mice were treated at the time points indicated (every 48 hr; according to ethically approved guidelines) with 40 mg·kg −1 probenecid (water soluble; ThermoFisher) or 20 mg·kg −1 AZ11645373 (Torcis) via the intranasal route in 50‐μl PBS.

Techniques: Infection

Journal: British Journal of Pharmacology

Article Title: Repurposing drugs targeting the P2X7 receptor to limit hyperinflammation and disease during influenza virus infection

doi: 10.1111/bph.14787

Figure Lengend Snippet: Administration of probenecid or AZ11645373 reduces hyperinflammation in the airways during HKx31 infection. Groups of C57BL/6 mice were infected intranasally with a high dose of HKx31 (10 5 PFU; n = 8 per group). Mice were treated intranasally with probenecid (40 mg·kg −1 ), AZ11645373 (20 mg·kg −1 ), or PBS on day 3 post‐infection, and 24 hr later, mice were killed. (a) Total numbers of leukocytes in BAL were determined by viable cell counts and (b–e) Ly6G + neutrophils, total CD11c + I‐A b low macrophages, CD11c + I‐A b high dendritic cells, and Ly6C + inflammatory macrophages in BAL were determined by flow cytometry. (f, i) Pro‐inflammatory cytokine levels were determined by ELISA or cytokine bead array in BAL fluid. (j) Viral loads in the lungs were measured by a standard plaque assay. Data presented are the means ± SEM from eight mice per group. * P < .05, significantly different from PBS; one‐way ANOVA

Article Snippet: Following infection, mice were treated at the time points indicated (every 48 hr; according to ethically approved guidelines) with 40 mg·kg −1 probenecid (water soluble; ThermoFisher) or 20 mg·kg −1 AZ11645373 (Torcis) via the intranasal route in 50‐μl PBS.

Techniques: Infection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Plaque Assay

Journal: British Journal of Pharmacology

Article Title: Repurposing drugs targeting the P2X7 receptor to limit hyperinflammation and disease during influenza virus infection

doi: 10.1111/bph.14787

Figure Lengend Snippet: Early treatment with probenecid or AZ11645373 results in improved efficacy. Groups of C57BL/6 mice were infected intranasally with (a, c) 10 5 PFU of HKx31 or (b, d) 50 PFU of PR8 ( n = 8 per group). Mice were treated intranasally with probenecid (40 mg kg −1 ), AZ11645373 (20 mg kg −1 ), or PBS commencing prior to the induction of severe disease (day 1 HKx31 or day 5 PR8) and every 48 hr thereafter (arrows). (a, b) Mice were weighed daily, and results are expressed as mean per cent weight change ± SEM. (c, d) Survival curves are shown. * P < .05 PBS, significantly different from AZ11645373; # P < .05, PBS significantly different from probenecid; Mantel–Cox log‐rank test

Article Snippet: Following infection, mice were treated at the time points indicated (every 48 hr; according to ethically approved guidelines) with 40 mg·kg −1 probenecid (water soluble; ThermoFisher) or 20 mg·kg −1 AZ11645373 (Torcis) via the intranasal route in 50‐μl PBS.

Techniques: Infection