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Updated: Nov 19, 2025
Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
Published on: April 19, 2017
Dendritic cells inclusion and cell-subset assessment improve flow-cytometry-based proliferation test in non-immediate
Ruben Fernandez-Santamaria1, Gador Bogas2, Francisca Palomares1
1Allergy Research Group, Instituto de Investigación Biomédica de Málaga-IBIMA, Málaga, Spain.
Insights
A new drug hypersensitivity test using drug-primed dendritic cells (DCs) significantly improves diagnosis. This flow-cytometry method enhances sensitivity for non-immediate drug hypersensitivity reactions (NIDHRs) by analyzing specific effector cells.
Area of Science:
- Immunology
- Cellular Biology
- Dermatology
Background:
- Non-immediate drug hypersensitivity reactions (NIDHRs) pose diagnostic challenges.
- Conventional lymphocyte transformation tests (LTTs) lack standardization and exhibit low sensitivity.
- Advancements in drug presentation by dendritic cells (DCs) and effector cell analysis show promise.
Purpose of the Study:
- To evaluate the added value of drug-primed DCs in LTT for NIDHRs.
- To assess the proliferative response of lymphocyte subpopulations using flow cytometry.
- To improve the diagnostic accuracy of in vitro tests for NIDHRs.
Main Methods:
- Patients with confirmed NIDHRs underwent conventional LTT (C-LTT) and drug-primed DCs LTT (dDC-LTT).
- Proliferative responses in T cells and other effector cells were analyzed using CFSE and flow cytometry.
- Specific effector cell subpopulations (e.g., Th1, NK cells) were identified for each clinical entity.
Main Results:
- Drug-primed DCs LTT (dDC-LTT) demonstrated significantly higher sensitivity (61.8%) than C-LTT (29.4%).
- Sensitivity varied by clinical entity: SJS-TEN (87.5% vs 62.5%), MPE (47.4% vs 15%), AGEP (80% vs 33%).
- Incorporating specific effector cell subpopulations achieved 100% sensitivity for SJS-TEN and AGEP, and 68.4% for MPE, with overall 87% sensitivity and 85% specificity.
Conclusions:
- Flow cytometry-based LTT utilizing drug-primed DCs enhances diagnostic sensitivity for NIDHRs.
- Focusing on specific effector cell subpopulations for each clinical entity further optimizes test performance.
- This approach offers a unique and improved in vitro cellular test for NIDHR diagnosis.
Background:
Lymphocyte transformation test (LTT) has been widely used to evaluate non-immediate drug hypersensitivity reactions (NIDHRs). However, the lack of standardization 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 (CFSE).
Results:
The C-LTT showed a significantly lower sensitivity (29.4%) compared with dDC-LTT (61.8%), which was confirmed analysing each particular clinical entity: SJS-TEN (62.5% vs 87.5%), MPE (15% vs 47.4%) and AGEP (33% vs 80%). When including the effector cell subpopulations involved in each clinical entity, CD3+ +CD4+ Th 1 or CD3+ +NK cells in SJS-TEN, CD3+ +CD4+ Th 1+NK cells in MPE and CD3+ +NK cells in AGEP, we could significantly increase the sensitivity of the in vitro test to 100%, 68.4% 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 focusing on effector cell subpopulations for each clinical entity significantly improved the drug-specific proliferative response in NIDHRs with a unique cellular in vitro test.
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