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Basophil Activation Test (basophil + activation_test)

Distribution by Scientific Domains

Medical Sciences	83%

Selected Abstracts

Basophil Activation Test and specific IgE measurements using a panel of recombinant natural rubber latex allergens to determine the latex allergen sensitization profile in children

PEDIATRIC ALLERGY AND IMMUNOLOGY, Issue 2 2006
María L. Sanz
There are no documented studies that describe natural rubber latex (NRL) sensitization in children with a history of surgical intervention but without any congenital malformation (urogenital anomalies, spina bifida, etc.), although some authors have studied NRL allergy in children without a history of surgical intervention. The aim of this work was to evaluate the sensitization profile to single NRL allergens in children without spina bifida and without repeated surgical interventions, by using different recombinant and natural latex allergens in two analytical techniques: specific serum immunoglobulin E (IgE) quantification and flow cytometry determination of activated basophils expressing CD63, after stimulating cells from patients with NRL allergens. A total of 23 patients and 10 healthy children were selected. Conjunctival and in-use NRL provocation tests were carried out, as well as specific IgE determination in all patients' and controls' sera with the recombinant NRL allergens: rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9 and rHev b 11 and with NRL (k82) using appropriate ImmunoCAPs. The Basophil Activation Test (BAT) was performed with whole latex extract and with the recombinant allergens rHev b 5 and rHev b 6.01, as well as with the natural allergen Hev b 6.02. The sensitivity and the specificity of NRL-specific IgE (k82) were 100%. Positive IgE responses to rHev b 5 were found in sera of 10 children, to rHev b 6.01 in 16 and for rHev b 6.02 in 15 children's sera. Specific IgE to rHev b 8 was found in four sera of the children. We only found significant differences in sensitization to rHev b 5 in children with two or more surgical interventions compared with the non-intervened group or those with only one intervention. Specific IgE in sera of children with latex-fruit syndrome recognized rHev b 6.02, but not to rHev b 11. The patients sensitized to Hev b 8, Hev b 9 and/or Hev b 11 were atopic. The four patients presenting a positive response to the NRL profilin Hev b 8 were allergic to pollen. The BAT against whole NRL extract was positive in 22 of 23 children; against rHev b 5 in 14 of the patients studied; against rHev b 6.01 in seven cases and against nHev b 6.02 in 19 children. In all the control subjects, the results using this technique were negative. If combined rHev b 5, rHev b 6.01 and nHev b 6.02 together, BAT could detect 20 of the 23 children with latex allergy. The combined use of ImmunoCAP with all the recombinant NRL allergens and BAT with rHev b 5, rHev b 6.01 and nHev b 6.02, enabled the identification of NRL allergy in 22 of 23 patients. There is a positive and significant correlation between sensitization to Hev b 5 and the number of interventions. BAT and allergen-specific IgE determination could be used as first-line in vitro diagnostic tests in patients with NRL allergy. [source]

Comparison of basophil activation tests using CD63 or CD203c expression in patients with insect venom allergy

ALLERGY, Issue 9 2006
B. Eberlein-König
Background:, Flow cytometric basophil activation tests have been developed as cellular tests for in vitro diagnosis of IgE-mediated reactions. Different activation markers (CD63 or CD203c) with distinct ways of regulation have been used after stimulation with various allergens. Objective:, It was the aim of the present study to compare basophil activation tests by measuring both CD63 and CD203c upregulation in patients with insect venom allergy. Materials and methods:, 43 patients with a history of insect venom anaphylaxis were examined. A careful allergy history was taken, and skin tests and determination of specific IgE-antibodies were performed. Basophil activation tests (BAT) using CD63 or CD203c expression were done after stimulation with different concentrations of bee and wasp venom extracts. 25 healthy subjects with negative history of insect venom allergy were studied as controls. Results:, The CD203c protocol showed a slightly higher sensitivity than the CD63 protocol (97% vs. 89%) with regard to patients' history. The magnitude of basophil response was higher with CD203c in comparison to CD63 for both insect venoms. Specificity was 100% for the CD63 protocol and 89% for the CD203c protocol with regard to controls with negative history and negative RAST. Conclusion:, These results support the reliability of basophil activation tests using either CD63 or CD203c as cellular tests in the in vitro diagnosis of patients with bee or wasp venom allergy with a slightly higher sensitivity for the CD203c protocol. [source]

P38 mitogen-activated protein kinase signal transduction in the diagnosis and follow up of immunotherapy of wasp venom allergy,

CYTOMETRY, Issue 5 2010
Marjoke M. Verweij
Abstract Background: P38 mitogen-activated protein kinase (MAPK) is known to govern IgE-mediated basophil activation. Intracellular phosphorylated p38 MAPK (Pp38 MAPK) in IgE-activated basophils can be quantified flow cytometrically. Objectives: To study whether Pp38 MAPK constitutes a potential novel read-out for flow-assisted diagnosis of hymenoptera venom allergy and to investigate whether this marker allows follow-up of successful venom immunotherapy (VIT). Methods: Fifty-two patients with documented wasp venom allergy and seven wasp-stung asymptomatic control individuals were enrolled. Wasp venom-induced basophil activation was analyzed flow cytometrically with anti-IgE, anti-CD63, and anti-Pp38 MAPK to assess their activation status before starting immunotherapy. To assess whether p38 MAPK constitutes a candidate marker for monitoring VIT, we repeated the basophil activation test (BAT) in 25 patients on the fifth day of a build-up immunotherapy. In addition, we investigated whether the Pp38 MAPK-based BAT could contribute in the decision of discontinuing VIT in a cross-sectional analysis in 13 patients receiving treatment for 3 years and 14 patients receiving treatment for 5 years. Results: Patients exhibited a dose-dependent basophil activation with phosphorylation of p38 MAPK and upregulation of downstream CD63. In contrast, stung controls demonstrated a dose-dependent but "abrogated" signal transduction in basophils with less and shorter duration of the phosphorylation of p38 MAPK and without subsequent upregulation of CD63. When repeated after 5 days of VIT and when investigated cross-sectionally after 3 years or 5 years of maintenance therapy, no effect of VIT on the phosphorylation of p38 MAPK was demonstrable. Conclusions: This study discloses that not only basophils from patients, but also from the stung control individuals, respond to wasp venom stimulation with phosphorylation of p38 MAPK, although to a lesser extend. No clear effect of VIT on the phosphorylation of p38 MAPK was shown. Thus, although p38 MAPK provides an additional tool in the diagnosis of wasp venom allergy, it does not contribute to the decision whether to stop successful VIT. © 2010 International Clinical Cytometry Society [source]

CD203c-based basophil activation test in allergy diagnosis: Characteristics and differences to CD63 upregulation,

CYTOMETRY, Issue 5 2010
Eva M. Sturm
Abstract Background: The basophil activation test (BAT) based on CD203c upregulation has been validated as a reliable tool for the diagnosis of IgE-mediated allergies. Nevertheless, CD203c-based BAT is hardly comparable with that of CD63-based tests, as the mechanisms of CD203c versus CD63 induction differ considerably. The aim of the present study was to identify potent influencing factors of the CD203c-based BAT and to emphasize differences between CD63 and CD203c detection. Methods: CD203c-based BAT was determined in 82 healthy controls and in 79 allergic patients. The effects of interleukin (IL)-3 and degranulation enhancing substances were investigated and compared with CD63 upregulation. Furthermore, the influence of different storage conditions and incubation times was evaluated and the impact of antiallergic drugs on the test results was assessed. Results: CD203c and CD63 expression was rapidly upregulated reaching a maximum after 20,30 min. Basophil CD203c upregulation assayed after storage times up to 48 h declined already after 4 h. IL-3 treatment increased CD203c and CD63 baseline levels and decreased basophil CD203c responses in a dose-dependent manner. In contrast, cytochalasin B and latrunculin B did not affect CD203c responses but decreased CD63-based BAT. Finally, therapeutic concentrations of dimetindene and desloratadine did not affect CD203c upregulation. Conclusion: CD203c-based basophil activation test should be performed preferentially within 4 h after taking the blood samples. Priming and degranulation-enhancing factors are not required for CD203c-based BAT. In contrast to skin testing, CD203c-based BAT can be performed in patients undergoing antiallergic treatment. © 2010 International Clinical Cytometry Society [source]

Usefulness of the basophil activation test (BAT) in the diagnosis of life-threatening drug anaphylaxis

ALLERGY, Issue 9 2010
P. García-Ortega
No abstract is available for this article. [source]

Usefulness of basophil activation test in diagnosis of occupational nonasthmatic eosinophilic bronchitis

ALLERGY, Issue 7 2010
G. Pala
No abstract is available for this article. [source]

Methylprednisolone-induced anaphylaxis: diagnosis by skin test and basophil activation test

ALLERGY, Issue 4 2010
B. Ben Said
No abstract is available for this article. [source]

Immediate allergic reaction to atropine in ophthalmic solution confirmed by basophil activation test

ALLERGY, Issue 9 2009
P. Cabrera-Freitag
No abstract is available for this article. [source]

Anaphylaxis to amidotrizoate proved by skin testing and flow cytometry-based basophil activation test

ALLERGY, Issue 3 2009
P. Dewachter
No abstract is available for this article. [source]

Analysis of basophil activation by flow cytometry in pediatric house dust mite allergy

PEDIATRIC ALLERGY AND IMMUNOLOGY, Issue 4 2008
Miguel González-Muñoz
Detection of allergen-induced basophil activation by flow cytometry has been shown to be a useful tool for allergy diagnosis. The aim of this study was to assess the potential of this technique for the diagnosis of pediatric house dust mite allergy. Quantification of total and specific IgE and basophil activation test were performed to evaluate mite allergic (n = 24), atopic (n = 23), and non-allergic children (n = 9). Allergen-induced basophil activation was detected as a CD63-upregulation. Receiver operating characteristics (ROC) curve analysis was performed to calculate the optimal cut-off value of activated basophils discriminating mite allergic and non-allergic children. ROC curve analysis yielded a threshold value of 18% activated basophils when mite-sensitized and atopic children were studied [area under the curve (AUC) = 0.99, 95% confidence interval (CI) = 0.97,1.01, p < 0.001] with a sensitivity and specificity of 96% for 16 ,g/ml mite extract. Analysis of the data obtained with 1.6 ,g/ml mite extract defined a cut-off value of 8% activated basophils (AUC = 0.96, 95% CI = 0.91,1.01; p < 0.001) with a sensitivity of 82% and specificity of 100%. Comparison between mite allergic and non-allergic children produced a cut-off of 8% activated basophils (AUC = 1.0) with 16 ,g/ml allergen extract and a sensitivity and specificity of 100%. The same threshold and specificity values were obtained with 1.6 ,g/ml extract (AUC = 97%, 95% CI = 0.92,1.02; p < 0.001) but sensitivity decreased to 83%. Two atopic children showed negative skin prick and basophil activation tests and high specific IgE (>43 kU/l) values for Dermatophagoides pteronyssinus allergen. They also showed positive prick (wheal diameter >1.0 cm) and basophil activation (>87%) tests and high specific IgE (>100 kU/l) with shrimp allergen. Shrimp sensitization was demonstrated by high levels of Pen a 1-specific IgE (>100 kU/l). Cross-reactivity between mite and shrimp was confirmed by fluorescence enzyme immunoassay (FEIA-CAP) inhibition study in these two cases. This study demonstrated that the analysis of allergen-induced CD63 upregulation by flow cytometry is a reliable tool for diagnosis of mite allergy in pediatric patients, with sensitivity similar to routine diagnostic tests and a higher specificity. Furthermore, this method can provide additional information in case of disagreement between in vivo and in vitro test results. [source]

Usefulness of basophil activation tests for the diagnosis of IgE-mediated allergy to quinolones

ALLERGY, Issue 4 2010
B. Ben Said
No abstract is available for this article. [source]

IgE-mediated allergy to pristinamycin: the value of skin tests and basophil activation tests

ALLERGY, Issue 11 2009
B. Bensaid
No abstract is available for this article. [source]

Comparison of basophil activation tests using CD63 or CD203c expression in patients with insect venom allergy

Analysis of basophil activation by flow cytometry in pediatric house dust mite allergy

Characterization of peach thaumatin-like proteins and their identification as major peach allergens

CLINICAL & EXPERIMENTAL ALLERGY, Issue 9 2010
A. Palacín
Summary Background Peach is the most important fruit related to food allergy in the Mediterranean area. Pru p 3, its lipid transfer protein, has been described as the principal allergen responsible for cross-reactivities with other foods and pollen and the severity of clinical symptoms. However, the involvement of other allergenic families cannot be ruled out. Thaumatin-like proteins (TLPs) have been described as food allergen in several fruits, such as apple, cherry, kiwi and banana, and pollen. Objective To identify members of the TLP family in peach fruit and to characterize putative allergens. Methods Through two-dimensional (2D) electrophoresis of peach extract and immunodetections with a pool of peach-allergic patients, IgE-binding spots were identified and the corresponding proteins purified and characterized as allergens by in vitro and in vivo assays. Three isoforms, belonging to the TLP family, were purified by different chromatographic systems and characterized by N -terminal amino acid sequences, molecular weight determination (MALDI) and enzymatic activity analysis (,-1,3-gluconase test and inhibition growth of fungi). In the same way, their IgE-binding capacity and allergenic activity were tested by ELISA assays, basophil activation tests and skin prick tests (SPT). Results Two peach-TLPs, Pru p 2.0101 and Pru p 2.0201, were identified as IgE-binding spots by 2D electrophoresis. Another peach-TLP, Pru p 2.0301, was cloned and produced as recombinant protein in a yeast system. The three isoforms were purified and characterized as TLPs by immunoblotting with anti-chestnut TLP antibodies and anti-plant N -asparagine complex glycan (anti-cross-reactive carbohydrate determinant). All of them showed ,-1,3-glucanase activity and inhibition of fungal growth. The three TLPs were recognized by around 50% of the sera from 31 patients analysed in ELISA experiments. All three gave a positive response to an SPT and/or in basophil activation experiments. Conclusion Three isoforms, belonging to the TLP family, were identified in peach as principal allergens. Their prevalence, observed in in vitro, ex vivo and in vivo analyses, suggests that they are important allergens and should therefore be included in the routine diagnosis of peach allergy, at least in the Mediterranean area. Cite this as: A. Palacín, L. Tordesillas, P. Gamboa, R. Sanchez-Monge, J. Cuesta-Herranz, M. L. Sanz, D. Barber, G. Salcedo and A. Díaz-Perales, Clinical & Experimental Allergy, 2010 (40) 1422,1430. [source]