Assessment of TSLP, IL 25 and IL 33 in patients with shrimp allergy

Background Shrimp allergy is a growing problem among the European population. TSLP, IL-25 and IL-33 are involved in the pathophysiology of allergic diseases, including asthma and atopic dermatitis, as they activate the Th2-dependent immune response. Methods Thirty-seven patients (18 male and 19 female) with a positive history of symptoms associated with shrimp consumption were selected. All patients had blood samples taken to assess the concentration of allergen-specific IgE (sIgE) to house dust mites (HDM) and shrimp (Singleplex, quantitative method with cut off value > 0,35 kAU/L) as well as the level of allergen components using the ImmunoCap ISAC method (Microarray test, semi-quantitative with cut off value > 0,3 ISU-E). The concentrations of TSLP, IL-25 and IL-33 in the patients’ blood serum was assessed using the ELISA method (Cusabio). Twenty patients with negative allergy history of allergic disease tests were included in the control group. Results Among the 37 shrimp-allergic patients, ImmunoCap ISAC was identified the presence of sIgE to the available shrimp allergen components in only 14 cases (37.8%). TSLP and IL25 levels were significantly higher in the study group. No statistically significant correlation was found between the concentration of analyzed alarmins and the concentration of sIgE level to shrimp or HDM between the study and control groups. No statistically significant correlation was found between poly-sensitization occurring in patients and levels of TSLP, IL-25 and IL-33 . Conclusion In shrimp-allergic patients, the concentrations of TSLP and IL-25 were significantly higher than in the control group (1.33 vs. 0.49 and 157 vs. 39.36, respectively). There was no correlation between the concentrations of TSLP, IL-25 and IL-33 and the concentration of sIgE in the patients or the number of allergen components that the patients were sensitized to. Trial registration: Bioethics Committee 147/2015, 11.03.2015.

Understanding Systemic and Local Inflammation Induced by Nasal Polyposis: Role of the Allergic Phenotype

Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by persistent symptoms associated to the development of nasal polyps. To this day, the molecular mechanisms involved are still not well defined. However, it has been suggested that a sustained inflammation as allergy is involved in its onset. In this exploratory study, the aim was to investigate the effect of the allergic status in the development of CRSwNP. To achieve this, we recruited 22 patients with CRSwNP and classified them in non-allergic and allergic using ImmunoCAP ISAC molecular diagnosis. Plasma samples were analyzed using liquid chromatography coupled to mass spectrometry (LC-MS). Subsequently, significant metabolites from plasma that were commercially available were then analyzed by targeted analysis in some nasal polyps. Additionally, nasal polyp and nasal mucosa samples were examined for eosinophils, neutrophils, CD3+ and CD11c+ cells, as well as collagen deposition and goblet cell hyperplasia. We found that 9 out of the 22 patients were sensitized to some aeroallergens (named as allergic CRSwNP). The other 13 patients had no sensitizations (non-allergic CRSwNP). Regarding metabolomics, bilirubin, cortisol, lysophosphatidylcholines (LPCs) 16:0, 18:0 and 20:4 and lysophosphatidylinositol (LPI) 20:4, which are usually related to a sustained allergic inflammation, were unexpectedly increased in plasma of non-allergic CRSwNP compared to allergic CRSwNP. LPC 16:0, LPC 18:0 and LPI 20:4 followed the same trend in nasal polyp as they did in plasma. Comparison of nasal polyps with nasal mucosa showed a significant increase in eosinophils (p < 0.001) and neutrophils (p < 0.01) in allergic CRSwNP. There were more eosinophils in polyps of non-allergic CRSwNP than in their nasal mucosa (p < 0.01). Polyps from non-allergic CRSwNP had less eosinophils than the polyps of allergic CRSwNP (p < 0.05) and reduced amounts of collagen compared to their nasal mucosa (p < 0.001). Our data suggests that there is a systemic inflammatory response associated to CRSwNP in the absence of allergy, which could be accountable for the nasal polyp development. Allergic CRSwNP presented a higher number of eosinophils in nasal polyps, suggesting that eosinophilia might be connected to the development of nasal polyps in this phenotype.

Clinical utility of immunological methods based on the singleplex and multiplex ImmunoCap systems for diagnosis of shrimp allergy

Background Levels of specific IgE (sIgE) against allergen components can be assessed using multiplex assays or with highly sensitive, quantitative methods. The aim of this study was to compare the sensitivity and specificity of different immunological methods for diagnosis of shrimp allergy. Methods Twenty patients with positive skin prick tests for frozen tiger shrimp were selected for further examination. Blood samples were taken to assess concentrations of sIgE against the house dust mites Dermatophagoides pteronyssinus and D. farinae, shrimp allergen extract, allergen components Der p 1, Der p 2 and Pan a 1 (ImmunoCap), and the ImmunoCap ISAC 112 panel. Results All patients had elevated levels of sIgE against shrimp and D pteronyssinus. Eight patients were sensitized to Pen m 1, three patients were sensitized to Pen m 2, and two patients were sensitized to Pen m 4 (ISAC). ImmunoCap ISAC detected shrimp sensitization in 50% of patients. There was a strong correlation between concentrations of sIgE against Pen m1 and Der p 10 detected by ImmunoCap. Conclusions The singleplex ImmunoCap system remains the reference diagnostic method, but in the case of shrimp allergy ImmunoCap ISAC provided better insight into patient allergen profiles.

A Retrospective Clinical Audit of the ImmunoCAP ISAC 112 for Multiplex Allergen Testing

<b><i>Introduction:</i></b> Complex cases of multiple allergies can be particularly difficult to diagnose using standard methods such as skin prick tests and assessment of a patient’s allergic history. Multiplex allergy testing may improve outcomes for allergy patients by avoiding misdiagnosis and providing reassurance. The ImmunoCAP Immuno Solid-Phase Allergen Chip (ISAC) 112 is a CE-marked, molecular, multiplex, allergy test that can test for IgE antibodies to 112 components from 51 allergen sources. However, its clinical utility is unknown and is difficult to estimate due to the complexity of the diagnostic pathway in which it is used. <b><i>Objective:</i></b> To assess how the ImmunoCAP ISAC 112 is currently being used in UK practice. The patient populations in which it may have the most benefit were examined, and the sequence of other tests implemented alongside ISAC was determined. <b><i>Methods:</i></b> A retrospective audit of 100 patient cases from 2 UK tertiary allergy clinics was performed. Fifty paediatric and fifty adult cases were selected for audit. The indications for ordering an ISAC test, the other tests used alongside ISAC, and changes in management actioned by the ISAC test were investigated. <b><i>Results:</i></b> 73.6% of paediatric and 78% of adult patients referred for an ISAC test were suspected to have multiple sensitizations. The sequence of testing varied greatly between cases, but 70% of adult and 98% of paediatric patients had at least one other investigation prior to an ISAC test. In most cases, ISAC testing confirmed clinical suspicion. <b><i>Conclusions:</i></b> A prospective research study is necessary to further investigate the clinical utility and cost-effectiveness of the ISAC. A UK national registry would be of great benefit but will require a large resource base.

DEFINITION OF SENSITIZATION TO POLLEN ALLERGENS WORMWOOD AND HAZEL IN PATIENTS WITH RESPIRATORY ALLERGIC DISEASES BY IMMUNOBLOT AND MULTIPLEX COMPONENT TEST

The aim of our study was to evaluate the diagnostic parameters of immunoblot and Immunocap ISAC methods to determine allergic sensitization to wormwood and hazel in patients with respiratory allergic diseases – allergic rhinitis and bronchial asthma. Materials and methods. In this study, 40 patients with bronchial asthma and / or allergic rhinitis were examined with two different methods of specific allergic diagnosis (in vitro). The study was open-ended, comparative. Quantitative determination of specific IgE in the serum was performed using the RIDA® AllergyScreen immunoblot method (R-Biopharm AG, Germany) on the basis of the private laboratory of LLC “Allergy-Immunological Center KPP”. Immunocap ISAC testing was performed at “Forpost Allergy and Immunology Clinic”. Results and Discussion. Among patients, allergic sensitization to wormwood was 27.5% (11 people) in the presence of specific IgE by the Rida AllergyScreen method, 25.0% (10 people) in the presence of specific IgE using ImmunoCAP ISAC; sensitization to hazel allergen was 27.5% (11 people) in the presence of specific IgE on Rida AllergyScreen, 30.0% (12 people) in the presence of specific IgE from ImmunoCAP ISAC. For the determination of specific IgE to wormwood allergen the immunoblot has a high sensitivity and negative predictive value (100.00% in both cases) compared to ISAC, a high specificity of 96.67% (95% CI: 82.78; 99.92), but a positive predicitive value was 90.91% (95% CI: 59.28; 98.57) and accuracy was 97.50% (95% CI: 86.84; 99.94). Conclusions. Immunoblot compared to ISAC for the determination of specific IgE to hazel allergen has a relatively high specificity and negative predictive value (92.86% and 89,66%), but the sensitivity and positive predicitive value were 75.00% (95 % CI: 42.81; 94.51) and 81.82% (95% CI: 53.23; 94.68), respectively, and the accuracy of the method was 87.50% (95% CI: 73.20; 95.81).

Comparative study of Allergy Explorer (ALEX) versus ImmunoCAP platforms

Objective. In this study, technical performance of the new multiplex ALEX test was compared with results from ImmunoCAP single tests (tIgE, sIgE) and the multiplex platform ImmunoCAP ISAC sIgE 112. Materials and methods. Eleven whole allergen extracts and corresponding allergen components from different allergen groups were used for the analysis of 64-66 patients’ sera by all three platforms. Results. For the whole allergens, 55% false negative results were obtained with the ALEX test comparing to the ImmunoCAP sIgE tests while for allergen components the ALEX test gives 33% false negative results when compared to ImmunoCAP sIgE test results. Additionally, the ALEX test is characterized by a low dynamic range - the platform demonstrated no results above 36 kUA/L for samples giving >100 kUA/L using ImmunoCAP Specific IgE tests in the analysis of sIgE response to the whole allergens. For the allergen components, ALEX showed no results above 38 kUA/L for samples of up to 150 kUA/L according to ImmunoCAP Specific IgE test results. Comparing to ImmunoCAP single plex tests, ALEX show low dynamic range and poor agreement in quantitative results for tIgE and sIgE both for whole allergens and allergen components, while in the comparison with ImmunoCAP ISAC sIgE 112 platform, the agreement is better, but the sensitivity and dynamic range are still low._ Conclusions. The ALEX test has some serious limitations in its performance comparing to both types of ImmunoCAP platforms.

Comparative study of Allergy Explorer (ALEX) versus ImmunoCAP platforms

Objective. In this study, technical performance of the new multiplex ALEX test was compared with results from ImmunoCAP single tests (tIgE, sIgE) and the multiplex platform ImmunoCAP ISAC sIgE 112. Materials and methods. Eleven whole allergen extracts and corresponding allergen components from different allergen groups were used for the analysis of 64-66 patients sera by all three platforms. Results. For the whole allergens, 55% false negative results were obtained with the ALEX test comparing to the ImmunoCAP sIgE tests while for allergen components the ALEX test gives 33% false negative results when compared to ImmunoCAP sIgE test results. Additionally, the ALEX test is characterized by a low dynamic range - the platform demonstrated no results above 36 kUA/L for samples giving 100 kUA/L using ImmunoCAP Specific IgE tests in the analysis of sIgE response to the whole allergens. For the allergen components, ALEX showed no results above 38 kUA/L for samples of up to 150 kUA/L according to ImmunoCAP Specific IgE test results. Comparing to ImmunoCAP single plex tests, ALEX show low dynamic range and poor agreement in quantitative results for tIgE and sIgE both for whole allergens and allergen components, while in the comparison with ImmunoCAP ISAC sIgE 112 platform, the agreement is better, but the sensitivity and dynamic range are still low._ Conclusions. The ALEX test has some serious limitations in its performance comparing to both types of ImmunoCAP platforms.