Murdoch University Research Repository

Welcome to the Murdoch University Research Repository

The Murdoch University Research Repository is an open access digital collection of research
created by Murdoch University staff, researchers and postgraduate students.

Learn more

Blister fluid as a cellular input for ex vivo diagnostics in drug-induced severe cutaneous adverse reactions improves sensitivity and explores immunopathogenesis

Awad, A., Mouhtouris, E., Nguyen-Robertson, C.V., Holmes, N., Chua, K.Y.L., Copaescu, A., James, F., Goh, M.S., Aung, A.K., Godfrey, D.I., Phillips, E.J., Gibson, A., Almeida, C.F. and Trubiano, J.A. (2021) Blister fluid as a cellular input for ex vivo diagnostics in drug-induced severe cutaneous adverse reactions improves sensitivity and explores immunopathogenesis. Journal of Allergy and Clinical Immunology: Global . In Press.

[img]
Preview
PDF (Pre-proof)
Download (1MB) | Preview
Free to read: https://doi.org/10.1016/j.jacig.2021.11.001
*No subscription required

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 standardisation and poor sensitivity. Modern approaches to testing include measurement of IFN-γ release from patient peripheral blood mononuclear cells (PBMC) stimulated with the suspected causative drug.

Objective

We sought to improve ex vivo diagnostics for drug-induced SCAR by comparing enzyme-linked immunospot (ELISpot) sensitivities and flow cytometry-based intracellular cytokine staining (ICS) and cellular composition of separate samples (PBMC or blister fluid cells (BFC)) from the same donor.

Methods

IFN-γ release ELISpot and flow cytometry analyses were performed on donor-matched PBMC and BFC samples from four SCAR patients with distinct drug-hypersensitivity.

Results

Immune responses to suspected drugs were detected in both PBMC and BFC samples of two donors (Case 1 in response to ceftriaxone and Case 4 to oxypurinol), with BFC eliciting stronger responses. For two other donors, only BFC samples showed a response to meloxicam (Case 2) or sulfamethoxazole and its 4-nitro metabolite (Case 3). Consistently, flow cytometry revealed a greater proportion of IFN-γ-secreting cells in the BFC compared to PBMC. BFC cells from Case 3 were also enriched for memory/activation/tissue-recruitment markers over PBMC.

Conclusion

Analysis of BFC samples for drug-hypersensitivity diagnostics offers a higher sensitivity for detecting positive responses compared to PBMC. This is consistent with recruitment (and enrichment) of cytokine-secreting cells with a memory/activated phenotype into blisters.

Item Type: Journal Article
Murdoch Affiliation(s): Institute for Immunology and Infectious Diseases
Publisher: Elsevier Inc. on behalf of the American Academy of Allergy, Asthma & Immunology.
Copyright: © 2021 Published by Elsevier Inc. on behalf of the American Academy of Allergy, Asthma & Immunology.
URI: http://researchrepository.murdoch.edu.au/id/eprint/63173
Item Control Page Item Control Page

Downloads

Downloads per month over past year