Clinical & translational immunology · 2025
A semi-automated ASC speck assay to evaluate pyrin inflammasome activation.
СтатьяНазвание и аннотация: Перевод в очереди
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Dai Pei, Skinner Oliver, Lin Xufeng, Telfser Aiden, Ruiz-Diaz Stephanie, Saldanha Rohit G, Frith Katie, Lin Ming-Wei, Preece Kahn, Gray Paul E, Pinzon-Charry Alberto, Sullivan Anna, Adelstein Stephen, Tong Winnie Wy, Parker Matthew Js, Girgis Laila, Wainstein Brynn, Ojami Samar, Deenick Elissa K, Goldstein Leonard D, Rogers Michael J, Phan Tri Giang
Аннотация
OBJECTIVE: To develop a rapid functional assay to validate variants of uncertain significance (VUS) in the MEFV gene.
METHODS: Overactivity of the pyrin inflammasome pathway and ASC speck oligomerisation in response to stimulation with low concentrations of Clostridium difficile toxin A was directly visualised by immunofluorescence microscopy. A semi-automated algorithm was developed to count cells and ASC specks.
RESULTS: The semi-automated ASC speck assay is able to discriminate between healthy controls and patients with familial Mediterranean fever (FMF) and pyrin inflammasome overactivity with high sensitivity. It is also able to discriminate pyrin inflammasome overactivity from other autoinflammatory disease controls with high specificity.
CONCLUSION: The semi-automated ASC speck assay may be a useful test to functionally validate VUS in the MEFV gene and screen for pyrin inflammasome overactivity.
METHODS: Overactivity of the pyrin inflammasome pathway and ASC speck oligomerisation in response to stimulation with low concentrations of Clostridium difficile toxin A was directly visualised by immunofluorescence microscopy. A semi-automated algorithm was developed to count cells and ASC specks.
RESULTS: The semi-automated ASC speck assay is able to discriminate between healthy controls and patients with familial Mediterranean fever (FMF) and pyrin inflammasome overactivity with high sensitivity. It is also able to discriminate pyrin inflammasome overactivity from other autoinflammatory disease controls with high specificity.
CONCLUSION: The semi-automated ASC speck assay may be a useful test to functionally validate VUS in the MEFV gene and screen for pyrin inflammasome overactivity.