- Oral presentation
- Open Access
Drug-specific in vitro release of IFN-gamma in patients with delayed cutaneuos drug hypersensitivity reactions
© Porebski and Bosak; licensee BioMed Central Ltd. 2015
- Published: 11 March 2015
- Flow Cytometry
- Peripheral Blood Mononuclear Cell
- Antiepileptic Drug
- Cytokine Secreting
- Cell Culture Supernatant
One of the challenges for management of drug hypersensitivity reactions (DHR) is to detect a culprit drug. Although measures of interferon IFN-gamma production by patients' drug-specific T cells have been the widely utilized cytokine assay, a systematic comparison of different methods has not yet been reported.
A total of 16 patients with clinically well-established maculopapular exanthema due to antiepileptic drugs hypersensitivity in remission state and 15 drug-exposed control donors without DHR were included to the study. Peripheral blood mononuclear cells of investigated individuals were isolated and cultured under defined conditions with drugs. IFN-gamma production was measured with electrochemiluminescence array assay and ELISA (cytokine level in cell culture supernatant), ELISpot (cytokine secreting cells), flow cytometry (intracellular staining in CD3+ CD4+ cells).
IFN-gamma production could be demonstrated in 13 of 16 patients using electrochemiluminescence assay (sensitivity 81%), in 8 of 16 patients using ELISA (sensitivity 50%), in 6 of 16 and 7 of 16 patients using ELISpot (sensitivity 46%) and flow cytometry (sensitivity 57%), respectively. The sensitivity of combined measurements of drug-specific IFN-gamma by ELISpot, ELISA and flow cytometry achieved 88%. Healthy controls showed negative drug-specific IFN-gamma production in contrast to individuals with a known sensitivity in all tested read-out systems. The assays demonstrated a test specificity of 100% (electrochemiluminescence), 93% (ELISA), 100% (ELISpot) and 100% (flow cytometry).
Analysis of drug-specific IFN-gamma production by means of different assays proved a useful and reliable approach for the in vitro detection of drug hypersensitivities in the investigated population. Electrochemiluminescence array assay offers distinct advantage over the other tested assays, including a greater sensitivity, but its availability is limited because of the costs. Also combining different assays may be a feasible approach to identify the causative drug of DHR.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.