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Improvement of lymphocyte proliferation assessment in non-immediate drug hypersensitivity reactions using flow-cytometry
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  • Ruben Fernandez-Santamaria,
  • Gador Bogas,
  • Francisca Palomares,
  • Maria Salas,
  • Tahia Fernandez,
  • Isabel Jimenez,
  • Esther Barrionuevo,
  • Inmaculada Doña,
  • Maria Torres,
  • Cristobalina Mayorga
Ruben Fernandez-Santamaria
IBIMA-Regional University Hospital of Malaga-UMA

Corresponding Author:rubenfernandezsantamaria@gmail.com

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Gador Bogas
IBIMA–Regional University Hospital of Malaga–UMA
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Francisca Palomares
IBIMA, Regional University Hospital of Málaga, UMA
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Maria Salas
IBIMA-Regional University Hospital of Malaga, UMA
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Tahia Fernandez
IBIMA-Regional University Hospital of Malaga, Research Laboratory-Allergy Unit Malaga, ES
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Isabel Jimenez
Instituto de Investigación Biomédica de Málaga-IBIMA
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Esther Barrionuevo
Hospital Universitario 12 de Octubre
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Inmaculada Doña
IBIMA-Regional University Hospital of Malaga, UMA
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Maria Torres
Regional University Hospital of Malaga-IBIMA-UMA-BIONAND-ARADyAL
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Cristobalina Mayorga
IBIMA-Regional University Hospital of Malaga
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Background. Lymphocyte transformation test (LTT) has been widely used to evaluate non-immediate drug hypersensitivity reactions (NIDHRs). However, the lack of standardisation and the low sensitivity have limited its routine diagnostic use. The drug presentation by dendritic cells (DCs) and the assessment of proliferation on effector cells have shown promising results. Flow-cytometry-based methods can help apply these improvements. We aimed to assess the added value of using drug-primed-DCs and the determination of the proliferative response of different lymphocyte subpopulations in NIDHRs. Methods. Patients with confirmed NIDHR were evaluated by both conventional (C-LTT) and with drug-primed-DCs LTT (dDC-LTT) analysing the proliferative response in T-cells and other effector cell subpopulations by using the fluorescent molecule, carboxyfluorescein diacetate succinimidyl ester. Results. The C-LTT showed a significantly lower sensitivity (33.3%) compared with dDC-LTT (65.2%), which was confirmed analysing each particular clinical entity: SJS-TEN (62.5% vs 87.5%), MPE (14.3% vs 41.7%), and AGEP (33% vs 80%). When including the effector cell subpopulations involved in each clinical entity, CD3++CD4+Th1 cells in SJS-TEN, CD3++CD4+Th1+NK cells in MPE, and CD3++NK cells in AGEP, we could significantly increase the sensitivity of the in vitro test to 100%, 66.6%, and 100%, respectively. With an overall sensitivity of 87% and 85% of specificity in NIDHR. Conclusions. The use of a flow-cytometry-based test, DCs as drug presenting cells, and focussing on effector cell subpopulations for each clinical entity significantly improved the drug-specific proliferative response in NIDHRs with a unique cellular in vitro test.
10 Jun 2020Submitted to Allergy
11 Jun 2020Submission Checks Completed
11 Jun 2020Assigned to Editor
13 Jun 2020Reviewer(s) Assigned
28 Jun 2020Review(s) Completed, Editorial Evaluation Pending
30 Jun 2020Editorial Decision: Revise Minor
11 Aug 20201st Revision Received
12 Aug 2020Submission Checks Completed
12 Aug 2020Assigned to Editor
14 Aug 2020Reviewer(s) Assigned
27 Aug 2020Review(s) Completed, Editorial Evaluation Pending
28 Aug 2020Editorial Decision: Revise Minor
05 Oct 20202nd Revision Received
06 Oct 2020Assigned to Editor
06 Oct 2020Submission Checks Completed
12 Oct 2020Reviewer(s) Assigned
25 Oct 2020Review(s) Completed, Editorial Evaluation Pending
25 Oct 2020Editorial Decision: Revise Minor
16 Nov 20203rd Revision Received
17 Nov 2020Submission Checks Completed
17 Nov 2020Assigned to Editor
27 Nov 2020Reviewer(s) Assigned
06 Dec 2020Review(s) Completed, Editorial Evaluation Pending
09 Dec 20204th Revision Received
10 Dec 2020Assigned to Editor
10 Dec 2020Submission Checks Completed
20 Dec 2020Reviewer(s) Assigned
04 Jan 2021Review(s) Completed, Editorial Evaluation Pending
04 Jan 2021Editorial Decision: Accept