Clinical implication of detecting sensitization to iodinated radiocontrast media in the basophil activation test by flow cytometry

2021 ◽  
Vol 66 (12) ◽  
pp. 747-754
Author(s):  
N. V. Bychkova ◽  
P. A. Selivanov ◽  
N. M. Kalinina
Keyword(s):  
Flow Cytometry ◽  
Immune Response ◽  
T Lymphocytes ◽  
Basophil Activation Test ◽  
Basophil Activation ◽  
Activation Test ◽  
Radiocontrast Media ◽  
Type 2 Immune Response

The use of iodinated radiocontrast media is necessary for visualization. A number of patients have adverse effects of various nature and severity when these drugs are administered. Routine allergy tests do not provide adequate diagnosis of reactions to drugs in this group. The aim of this work is to assess the capabilities of the basophil activation test to confirm sensitization to non-ionic iodinated radiocontrast media, as well as to select a safe alternative drug in patients with a burdened history. Basophil activation test by flow cytometry was performed in 184 patients The Nikiforov Russian Centre of Emergency and Radiation Medicine» EMERCOM of Russia and 32 volunteers using ultravist, omnipack, and optiray. The presence of sensitization was assessed based on the basophil activation index, as well as spontaneous and anti-IgE antibody-induced activation of basophils and the population of T-lymphocytes type 2 immune response. The volunteers showed no sensitization to iodinated radiocontrast media. In patients with a medium degree of hypersensitivity reaction in vivo, in vitro sensitization to drugs was detected 4 times more often than in patients with a mild degree (51% versus 13.5%). In patients with systemic reactions to the administration of a known drug, in vitro sensitization was confirmed in 86% of cases, while the frequency of detection of sensitization to drugs did not differ. Spontaneous activation of basophils in patients and type 2 T-lymphocytes were 2 times higher than in volunteers. Patients were more likely to have low (less than 30%) activation of basophils for anti-IgE antibodies. The specificity of the basophil activation test with iodinated radiocontrast media was 100% with a sensitivity of 94.1%. Most patients were able to select a non-sensitizing contrast. Inclusion in the algorithm of spontaneous and anti-IgE antibody-induced activation of basophils and a population of T-lymphocytes type 2 immune response will allow the doctor to carry out a personalized approach to the management of patients with a burdened history.

scholarly journals Clinical and immunological characteristics of severe bronchial asthma with Aspergillus sensitization

2021 ◽  
Vol 5 (1) ◽  
pp. 10-16
Author(s):  
Ya.I. Kozlova ◽  
◽  
E.V. Frolova ◽  
A.E. Uchevatkina ◽  
L.V. Filippova ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Bronchial Asthma ◽  
Stimulation Index ◽  
Basophil Activation Test ◽  
Specific Ige ◽  
Forced Expiratory Volume ◽  
Basophil Activation ◽  
Activation Test ◽  
To Receive

Aim: to assess the clinical and immunological characteristics of patients with severe bronchial asthma (BA) with Aspergillus sensitization and to study the possibilities of applying basophil activation test (BAT) using flow cytometry to detect mycogenic sensitization. Patients and Methods: 93 patients with severe BA were examined. Therapy had 4 or 5 steps according to GINA 2019 recommendations. TARC (thymus and activation-regulated chemokine) content, the number of eosinophils, the levels of total IgE and specific IgE to 10 allergens were determined in blood serum by enzyme immunoassay. BAT with Aspergillus fumigatus allergen was performed in vitro using flow cytometry. Results: Aspergillus sensitization was detected in 33 (35.4%) patients with severe BA. In patients with BA and Aspergillus sensitization, the disease course was uncontrolled, and patients in this group were credibly more likely to receive oral glucocorticosteroids. The number of basophils (activated by the A. fumigatus allergen) and the stimulation index in patients with BA and Aspergillus sensitization were significantly higher than in patients with BA (9.9 (6.0–24.0) % vs. 3.6 (2.0–5.4) %; (p=0.000) and 4.25 (2.49–9.30) vs. 0.94 (0.75–1.16); (p=0.000)). Significant differences in TARC content were obtained in the groups of patients with severe BA and Aspergillus sensitization and patients with BA (625.0 (418.4–875.0) pg/mg versus 406.0 (210.0–561.0) pg/mg; p=0.001). A negative correlation was determined between TARC levels and a decrease in forced expiratory volume in 1 second (FEV1) (r=-0.70, p<0.05), and between positive correlation and absolute eosinophil count (r=0.81, p<0.05) and level of specific IgE to Aspergillus (r=0.36, p<0.05). Conclusion: Aspergillus sensitization is associated with an uncontrolled BA course. An additional method for diagnosing mycogenic sensitization is the BAT. The TARC concentration can serve as a biomarker of an active inflammatory response. KEYWORDS: Aspergillus spp., severe bronchial asthma, mycogenic sensitization, basophil activation test, TARC. FOR CITATION: Kozlova Ya.I., Frolova E.V., Uchevatkina A.E. et al. Clinical and immunological characteristics of severe bronchial asthma with Aspergillus sensitization. Russian Medical Inquiry. 2021;5(1):10–16. DOI: 10.32364/2587-6821-2021-5-1-10-16.

Immediate allergic reaction following intra-articular injection of ioxaglate confirmed by skin testing and flow cytometry-based basophil activation test

2010 ◽  
Vol 74 (2) ◽  
pp. e27-e29 ◽  
Author(s):  
Pascale Dewachter ◽  
Samuel Castro ◽  
Frédéric Zeitoun ◽  
Sylvie Chollet-Martin ◽  
Laurence Gaillanne ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Allergic Reaction ◽  
Skin Testing ◽  
Basophil Activation Test ◽  
Basophil Activation ◽  
Activation Test

scholarly journals Basophil activation test by flow cytometry: Present and future applications in allergology

2008 ◽  
Vol 74B (4) ◽  
pp. 201-210 ◽  
Author(s):  
D. G. Ebo ◽  
C. H. Bridts ◽  
M. M. Hagendorens ◽  
N. E. Aerts ◽  
L. S. De Clerck ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Basophil Activation Test ◽  
Basophil Activation ◽  
Activation Test

scholarly journals Application of Basophil Activation Test with Anthraxin for Laboratory ( in vitro) Diagnostics of Anthrax

2012 ◽  
pp. 86-88 ◽  
Author(s):  
A. N. Kulichenko ◽  
E. L. Rakitina ◽  
D. G. Ponomarenko ◽  
O. V. Logvinenko ◽  
A. G. Ryazanova
Keyword(s):  
Basophil Activation Test ◽  
Basophil Activation ◽  
Activation Test

scholarly journals Methylprednisolone-induced Anaphylaxis Diagnosed by Intradermal Skin Test Not Basophil Activation Test: A Case Report

2020 ◽  
Author(s):  
Hitomi Amano ◽  
Yoshiro Kitagawa ◽  
Taichiro Muto ◽  
Akihisa Okumura ◽  
Hideyuki Iwayama
Keyword(s):  
Skin Test ◽  
Incubation Time ◽  
Sodium Phosphate ◽  
Sodium Succinate ◽  
Basophil Activation Test ◽  
Basophil Activation ◽  
Diagnostic Strategy ◽  
Activation Test

Abstract BackgroundAnaphylaxis is a severe systemic allergic reaction. Glucocorticoids rarely induce anaphylaxis. Determination of allergens includes the in vivo skin prick test (SPT) and intradermal skin test (IDST) and the in vitro basophil activation test (BAT). However, the usefulness of BAT in determining drug allergens has not been adequately studied.Case presentation A 10-year-old boy was admitted to our hospital because of fever and arthralgia for 2 weeks. He had not been treated with glucocorticoids. According to the laboratory tests and imaging studies, he was suspected to have bacterial myositis and was treated with ceftriaxone. However, his symptoms persisted for more than 2 weeks. With a suspicion of autoinflammatory arthritis, we planned methylprednisolone (mPSL) sodium succinate (MPS) during pulse therapy (30 mg/kg). Fifteen minutes after the injection of mPSL, he had wheezing and generalized wheal formation with decreased oxygenation. The administration of mPSL was discontinued because anaphylaxis was suspected. Thirty minutes after the administration of oxygen and oral olopatadine, the anaphylactic symptoms resolved. One month after discharge, SPT, IDST, and BAT were performed under the administration of oral prednisolone. The SPTs for MPS, hydrocortisone sodium succinate (HCS) and prednisolone sodium succinate (PSS) were negative. The IDST for MPS was positive. Moreover, the IDSTs for HCS and PSS were positive, whereas those for dexamethasone sodium phosphate and betamethasone sodium phosphate were negative. The BAT for MPS was negative at 1.0% and 1.9% after an incubation time of 1 hour and 24 hours, respectively, although the BAT for histamine as positive control was 60.4% and 18.3% after an incubation time of 1 hour and 24 hours, respectively. The BATs for HCS and PSS were negative. Therefore, we diagnosed as anaphylaxis secondary to the succinate ester in MPS.ConclusionsIn this case, IDST was useful for the diagnosis of MPS-induced anaphylaxis, whereas BAT was negative. This highlighted the need to choose the appropriate procedure in the diagnosis of steroid-induced anaphylaxis. The results in our patient suggested that BAT may be considered when IDST and SPT are negative. Further studies are necessary to clarify the diagnostic strategy for steroid-induced anaphylaxis.

Developing an in Vitro Functional Model of Allergic Transfusion Reactions

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3426-3426
Author(s):  
William Savage ◽  
Aaron A. R. Tobian ◽  
Jessica Savage ◽  
Chris Thoburn ◽  
Paul M. Ness ◽  
...  
Keyword(s):  
Basophil Activation Test ◽  
Positive Control ◽  
Basophil Activation ◽  
Clinical Severity ◽  
Blue Staining ◽  
Activation Test ◽  
And Control ◽  
Transfusion Reactions ◽  
Positive Controls

Abstract Abstract 3426 Background: Allergic transfusion reactions (ATRs) occur in ∼2–3% of platelet transfusions, but their etiology remains largely unclear. Both donor and recipient factors have been shown to contribute to ATR risk. An in vitro model of ATRs would enable mechanistic studies. Objective: To develop a basophil activation test as a model of ATRs to apheresis platelets (AP) by measuring histamine release (HR) and leukotriene C4 (LTC4) production after exposing basophils to AP supernatants. Methods: Basophil-enriched suspensions (∼5–15% basophils) from healthy donors were prepared using double-Percoll density centrifugation. After washing and counting basophil numbers using Alcian blue staining, cells were resuspended in buffer containing 2 ng/mL IL-3 and 5mM Ca++/Mg++ such that 20–30×104 basophils were added in 0.025mL aliquots. IL-3 is a potent basophil priming agent and was used to model the increased recipient susceptibility to allergic reactions that has been observed clinically. Reaction tubes (1.5 mL) were set-up containing 0.025mL of 2× (twice the final conc.) of ATR/control supernatant (final titer 1:2–1:8), negative (buffer alone), and positive control reagents (100 ng/mL anti-IgE antibody, 50 ng/mL C5a, 1 μM FMLP). Cells and reaction tubes were pre-warmed separately to 37°C for 15' before aliquoting 0.025mL cell suspension to reaction tubes for a final volume of 0.050mL. After 30′ incubation at 37°C, 0.950mL of cold buffer (without IL-3 or Ca++/Mg++) was added to each reaction tube. Tubes were centrifuged 10" at ∼300g (4°C) before removing 0.900mL for histamine detection by automated fluorimetry and LTC4 detection by ELISA. AP supernatants that were (n=10) or were not (n=10) associated with ATRs were tested. HR values were determined as a percentage of the total histamine content by lysis with perchloric acid (1.6%). Results used in analysis are net HR or LTC4 production (AP sample minus IL-3 primed negative control) normalized to HR or LTC4 production observed with positive control. Results: IL-3 priming of basophils was necessary in order to evoke a response after exposure to AP supernatant. HR in response to AP incubation with unprimed basophils was <3% of positive controls. Maximal HR and LTC4 production in response to AP supernatants occurred at a final titer of 1:4 using IL-3 primed basophils. HR and LTC4 production within individual reactions were correlated (Spearman rho=0.83, P<0.0001). Mean net HR among all AP supernatant samples was 55±28%, 60±49%, and 53±29% of c5a, anti-IgE, and FMLP positive controls, respectively. LTC4 production was 19±26%, 14±22%, and 15±20% of c5a, anti-IgE, and FMLP positive controls. No statistically significant differences were observed in HR or LTC4 production when ATR and control groups were compared (P>0.1). ATRs were stratified into clinical categories of mild (pruritus/urticaria, n=3), moderate (angioedema/dyspnea, n=5), and severe (any symptom with hypotension, n=2). There were no differences or trends in HR and LTC4 production among the categories (P>0.2). There was no correlation between pre/post transfusion tryptase elevations in recipient plasma (n=4) and HR or LTC4 production. HR and LTC4 production was similar among blood types of AP products (A: n=12, O: n=5, AB: n=3), (P>0.7). We previously reported that concentrations of C5a are higher in AP products associated with ATRs than controls. C5a content in AP supernatant modestly correlated with basophil HR release (rho: 0.47, P=0.04), but less with LTC4 production (rho: 0.27, P=0.3). Summary: An in vitro basophil activation test that measures HR and LTC4 production was developed to study ATRs. We find that 1) IL-3 priming of basophils was necessary to elicit responses to AP supernatants; 2) LTC4 and HR are correlated within each reaction; 3) there are no differences in HR and LTC4 production between ATR and control AP supernatants were observed, regardless of clinical ATR severity; and 4) HR is associated with C5a content in the AP supernatant. Conclusions: The observations of 1) an overall requirement for basophil priming and 2) the lack of correlation between the clinical severity of ATRs and basophil HR and LTC4 release implicate recipient atopic priming as a risk factor for ATRs. However, the association of AP C5a content with HR suggests a donor/product role in ATRs, as well. Studies focusing on the nature of recipient susceptibility may further elucidate mechanisms of ATRs. Disclosures: No relevant conflicts of interest to declare.

Reliability of Basophil Activation Test Using CD203c Expression in Diagnosis of Pollen Allergy

2011 ◽  
Vol 25 (6) ◽  
pp. e225-e231 ◽  
Author(s):  
Seçil Kepil Özdemir ◽  
Deniz Güloğlu ◽  
Betül A. Sin ◽  
Atilla H. Elhan ◽  
Aydan İkincioğulları ◽  
...  
Keyword(s):  
Grass Pollen ◽  
Immunoglobulin E ◽  
Pollen Allergy ◽  
Basophil Activation Test ◽  
Basophil Activation ◽  
Pollen Extract ◽  
Activation Test ◽  
Cutoff Values ◽  
Phl P 5

Background CD203c is a basophil surface marker and its expression is rapidly up-regulated after cross-linking of high-affinity immunoglobulin E (IgE) receptor (FcepsilonR1) by an allergen. CD203c basophil activation tests have been studied for the in vitro diagnosis of several allergic conditions. However, there is limited data about its diagnostic usefulness. The optimum allergen concentrations for stimulation and allergen specific cutoff values remain unknown for a number of allergens. This study was designed to investigate the efficacy of basophil activation test via CD203c in the diagnosis of pollen allergy. Methods The CD203c basophil activation was determined in 31 allergic rhinitis patients with pollen allergy and 9 healthy nonatopic controls during the off-season. CD203c expression was evaluated using three-color staining protocol by flow cytometry. Results After an in vitro stimulation with grass pollen extract, the CD203c assay clearly discriminated pollen-allergic patients from controls (p < 0.001). A dose-dependent increase in the percentages of CD203c-activated basophils was shown in rhinitis patients with pollen allergy (p < 0.001). The sensitivity and specificity was 100% and optimal cutoff values were 14.05 and 10.05% with 45.1 and 4.5 μg/mL Phl p 5 stimulation, respectively. Although the specificity was also 100%, the sensitivity was 93 and 87% and the cutoff values were 5.40 and 5.35% with 4.5 x 10– 4 and 4.5 x 10– 5 micrograms/mL Phl p 5 stimulation, respectively. Conclusion The CD203c basophil activation test seems to be a reliable tool in the diagnosis of grass pollen allergy. It could be used when conventional diagnostic tests fail or can not be performed.

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