In-vivo and ex-vivo tests for culprit drugs identification in severe cutaneous adverse drugs reactions.

Drug causality assessment in severe cutaneous adverse reactions (SCARs) remains challenging. We investigated the usefulness of in-vivo drug patch tests (PT), ex-vivo interferon (IFN)-γ enzyme-linked immunospot (ELISpot) assay, and lymphocyte transformation test (LTT) in 30 SCARs patients within the past 36 months. Drug PT yielded a 20% positivity rate (n = 6), while IFN-γ ELISpot and LTT showed positive rates of 56.67% (n = 17) and 41.38% (n = 12), respectively. Combining the three tests resulted in an overall positive rate of 66.67% (n = 20) of cases. IFN-γ ELISpot offered additional positivity, especially with oxypurinol. Employing a combined diagnostic approach may enhance the chances of obtaining a positive result.

Acute urticaria alone after CoronaVac COVID-19 vaccination should not be a contraindication for revaccination.

Patients who develop an immediate allergic reaction within the first 4 h of COVID-19 vaccine injection are recommended not to receive the same vaccine again. This recommendation mainly focuses on the mRNA and adenoviral vector COVID-19 vaccines, but data for whole virus vaccines are unknown. We report seven patients who developed an immediate reaction within 4 h (six had generalized urticaria, one had localized urticaria) after the first vaccination with CoronaVac, the inactivated SARS-CoV-2 vaccine. The results of skin tests and basophil activation tests suggested that spike peptides play a role in exacerbating urticaria in some patients. However, all subjects who developed urticaria within 4 h after CoronaVac vaccination were successfully revaccinated without graded challenge, although recurrent urticaria was common. This preliminary result indicates that acute urticaria alone should not be a contraindication for the second dose of CoronaVac if the supply of alternative vaccines is limited.

The measurement of drug-induced interferon γ-releasing cells and lymphocyte proliferation in severe cutaneous adverse reactions.

BACKGROUND: The lymphocyte transformation test (LTT) is a standard laboratory method to identify culprit drugs in patients with a history of drug-induced non-immediate hypersensitivity and is mainly performed during the recovery phase. The measurement of drug-specific interferon γ (IFN-γ)-releasing cells has been introduced to confirm culprit drugs, even during the acute phase of drug allergy. OBJECTIVES: This study aimed to evaluate the capability of the enzyme-linked immunospot assay (ELISpot) to detect drug-specific IFN-γ-releasing cells during the acute phase and the capability of LTT to identify culprit drugs during the recovery phase in patients presenting with severe cutaneous adverse reactions (SCARs). METHODS: Peripheral blood mononuclear cells (PBMCs) from 23 SCAR patients were collected during the acute and recovery phases and assayed for drug-specific IFN-γ-releasing cells and lymphocyte proliferation, respectively. RESULTS: Drug-specific IFN-γ-releasing cells were detectable in 73.9% of SCAR subjects (55.6% and 85.7% in patients who were and were not taking systemic steroids, respectively), whereas LTT results were positive in 52.2% of SCAR subjects. The frequencies of drug-specific IFN-γ-releasing cells were significantly higher in patients with positive LTT than in those with negative LTT (260.1 ± 110.0 and 46.6 ± 20.7 cells/106 PBMCs, P = 0.01). A significant correlation between the results of the IFN-γ ELISpot assay and LTT was demonstrated (r = 0.65, P value <0.01). CONCLUSION: The IFN-γ ELISpot assay could be a useful tool to identify culprit drugs in SCAR patients when culprit drug identification is urgently needed during the acute phase of drug allergy.

In vitro test to confirm diagnosis of allopurinol-induced severe cutaneous adverse reactions.

BACKGROUND: Allopurinol is a frequent cause of severe cutaneous adverse reactions (SCARs), such as drug reaction with eosinophilia and systemic symptoms (DRESS), Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). The reactions can potentially be fatal. As drug rechallenge in patients with a history of drug-induced SCARs is contraindicated, in vitro testing may have a diagnostic role as a confirmation test. OBJECTIVES: To study the diagnostic value of interferon (IFN)-γ enzyme-linked immunospot (ELISpot) assay as a confirmatory test in patients with a history of allopurinol-induced SCARs. METHODS: Peripheral blood mononuclear cells (PBMCs) from 24 patients with a history of allopurinol-induced SCAR (13 DRESS, 11 SJS/TEN) and 21 control subjects were incubated with allopurinol or oxypurinol in the presence or absence of antiprogrammed death ligand 1 antibody (anti-PD-L1). The numbers of IFN-γ-releasing cells after stimulation in each group were subsequently measured with ELISpot. RESULTS: The numbers of IFN-γ-releasing cells in allopurinol-allergic subjects were significantly higher than in control subjects when stimulating PBMCs with oxypurinol 100 µg mL-1 , especially when adding anti-PD-L1 supplementation. According to the receiver operating characteristic curve results, the optimal discriminatory power of IFN-γ ELISpot in confirming diagnosis of allopurinol-induced SCARs can be obtained using 16 spot-forming cells per 106 PBMCs as a cut-off value upon oxypurinol/anti-PD-L1 stimulation (79·2% sensitivity and 95·2% specificity). CONCLUSIONS: The measurement of oxypurinol/anti-PD-L1-inducing IFN-γ-releasing cells yields a high diagnostic value in distinguishing between allopurinol-allergic and control subjects. This technique is beneficial in confirming diagnosis of allopurinol-induced SCARs in patients whose reaction develops while taking multiple drugs.

The values of nasal provocation test and basophil activation test in the different patterns of ASA/NSAID hypersensitivity

Background: The oral provocation test (OPT) is the current gold standard to diagnose aspirin hypersensitivity syndrome although it is time-consuming and contains some systemic risks. Other reliable methods with lower side effects and shorter test duration are being investigated. Objective: The purpose of this study was to evaluate the efficacy of the nasal provocation test (NPT) and the basophil activation test (BAT) in the diagnosis of different subtypes of aspirin sensitivity. Methods: Thirty aspirin sensitivity patients with cutaneous and respiratory manifestations underwent NPT and BAT with lysine-ASA. NPT result was interpreted as recommended in EAACI/GA2LEN guidelines and receiver operating characteristic analysis of BAT was performed by using 15 NSAIDs tolerant volunteers as a control group. Results: NPT was positive in 60% (18/30) of patients and BAT was positive in 76.7% (23/30) of patients. The incubation of basophils with 0.31mg/ml of lysine-aspirin and using 4.6% activated basophils gives the best predictive values to diagnose aspirin sensitivity. The combination of both tests yielded positive results in 80% and 93.3% of aspirin-induced cutaneous and respiratory patterns. The agreement between NPT and BAT results was 63.3%. Conclusions: NPT and BAT are beneficial to detect patients with aspirin sensitivity. The combination of both tests have additional diagnostic values; less time-consuming than OPT and their complications are negligible. A reliable alternative method with minimum side effects is needed to diagnose aspirin sensitivity in suspected patients who have contraindications for OPT(AU)

The values of nasal provocation test and basophil activation test in the different patterns of ASA/NSAID hypersensitivity.

BACKGROUND: The oral provocation test (OPT) is the current gold standard to diagnose aspirin hypersensitivity syndrome although it is time-consuming and contains some systemic risks. Other reliable methods with lower side effects and shorter test duration are being investigated. OBJECTIVE: The purpose of this study was to evaluate the efficacy of the nasal provocation test (NPT) and the basophil activation test (BAT) in the diagnosis of different subtypes of aspirin sensitivity. METHODS: Thirty aspirin sensitivity patients with cutaneous and respiratory manifestations underwent NPT and BAT with lysine-ASA. NPT result was interpreted as recommended in EAACI/GA2LEN guidelines and receiver operating characteristic analysis of BAT was performed by using 15 NSAIDs tolerant volunteers as a control group. RESULTS: NPT was positive in 60% (18/30) of patients and BAT was positive in 76.7% (23/30) of patients. The incubation of basophils with 0.31 mg/ml of lysine-aspirin and using 4.6% activated basophils gives the best predictive values to diagnose aspirin sensitivity. The combination of both tests yielded positive results in 80% and 93.3% of aspirin-induced cutaneous and respiratory patterns. The agreement between NPT and BAT results was 63.3%. CONCLUSIONS: NPT and BAT are beneficial to detect patients with aspirin sensitivity. The combination of both tests have additional diagnostic values; less time-consuming than OPT and their complications are negligible. A reliable alternative method with minimum side effects is needed to diagnose aspirin sensitivity in suspected patients who have contraindications for OPT.