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Title: Blister fluid as a cellular input for ex vivo diagnostics in drug-induced severe cutaneous adverse reactions improves sensitivity and explores immunopathogenesis
Austin Authors: BPharm, Andrew Awad;Mouhtouris, Effie;Nguyen-Robertson, Catriona Vi;Holmes, Natasha E ;Chua, Kyra Y.L. ;Copaescu, Ana ;James, Fiona;Goh, Michelle S.;Aung, Ar Kar.;Godfrey, Dale I.;Philips, Elizabeth J.;Andrew, Gibson;Almeida, Catarina F.;Trubiano, Jason 
Affiliation: Infectious Diseases
Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Melbourne, Australia
Department of Dermatology, Alfred Health, Melbourne, Australia
Department of General Medicine, Alfred Health, Melbourne, Australia
The Institute for Immunology and Infectious Diseases, Murdoch University, Murdoch, Australia
Medicine (University of Melbourne)
Issue Date: 30-Nov-2022
Publication information: Journal of Allergy and Clinical Immunology: Global 2022; 1 (1): 16-21
Abstract: Background Drug-induced severe cutaneous adverse reactions (SCARs) are presumed T-cell–mediated hypersensitivities associated with significant morbidity and mortality. Traditional in vivo testing methods, such as patch or intradermal testing, are limited by a lack of standardization and poor sensitivity. Modern approaches to testing include measurement of IFN-γ release from patient PBMCs stimulated with the suspected causative drug. Objective We sought to improve ex vivo diagnostics for drug-induced SCARs by comparing enzyme-linked immunospot (ELISpot) sensitivities and flow cytometry–based intracellular cytokine staining and determination of the cellular composition of separate samples (PBMCs or blister fluid cells [BFCs]) from the same donor. Methods ELISpot and flow cytometry analyses of IFN-γ release were performed on donor-matched PBMC and BFC samples from 4 patients with SCARs with distinct drug hypersensitivity. Results Immune responses to suspected drugs were detected in both the PBMC and BFC samples of 2 donors (donor patient 1 in response to ceftriaxone and case patient 4 in response to oxypurinol), with BFCs eliciting stronger responses. For the other 2 donors, only BFC samples showed a response to meloxicam (case patient 2) or sulfamethoxazole and its 4-nitro metabolite (case patient 3). Consistently, flow cytometry revealed a greater proportion of IFN-γ–secreting cells in the BFCs than in the PBMCs. The BFCs from case patient 3 were also enriched for memory, activation, and/or tissue recruitment markers over the PBMCs. Conclusion Analysis of BFC samples for drug hypersensitivity diagnostics offers a higher sensitivity for detecting positive responses than does analysis of PBMC samples. This is consistent with recruitment (and enrichment) of cytokine-secreting cells with a memory/activated phenotype into blisters.
DOI: 10.1016/j.jacig.2021.11.001
ORCID: 0000-0003-4604-1062
Journal: Journal of Allergy and Clinical Immunology: Global
Type: Journal Article
Subjects: Severe cutaneous drug reactions
ex vivo assays
Appears in Collections:Journal articles

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