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PubMed2004

In vitro testing to diagnose venom allergy and monitor immunotherapy: a placebo-controlled, crossover trial.

BACKGROUND: In people with a history of sting allergy, only prior reaction severity and older age are known to predict subsequent reaction risk. Furthermore, no diagnostic test other than a deliberate sting challenge has been found to identify people in whom venom immunotherapy (VIT) has been unsuccessful. OBJECTIVE: We aimed to assess the utility of a number of in vitro tests to diagnose venom allergy and to monitor immunotherapy. METHODS: During a double-blind randomized placebo-controlled crossover trial of Myrmecia pilosula ant VIT the following venom-specific tests were performed at enrolment, and at completion of treatment prior to a diagnostic sting challenge; leucocyte stimulation index (SI), IL-4 production, IgE RAST, histamine release test (HRT), leukotriene release test (LRT) and basophil activation test (BAT). Intradermal venom skin testing (VST) was also performed at trial entry. RESULTS: Only VST and HRT identified those at risk of sting anaphylaxis in the placebo group. Although IgE RAST, leucocyte SI and IL-4 production, LRT and BAT all correlated well with intradermal VSTs, they did not predict sting challenge outcome. After successful VIT, venom-induced leucocyte IL-4 production tended to fall, whereas IgE RAST increased and a natural decline in HRT reactivity was reversed. A confounding seasonal affect on laboratory results was suspected. CONCLUSION: The HRT warrants further assessment for diagnosis of venom allergy. Uninformative performance of the commercially available LRT and BAT tests may be due to pre-incubation with IL-3. None of the tests evaluated appear to be reliable markers of successful VIT.

Animals↗

PubMed1990

Basophil histamine release in the diagnosis of house dust mite and dander allergy of asthmatic children. Comparison between prick test, RAST, basophil histamine release and bronchial provocation.

The aim of the study is to compare the glass fibre-based basophil histamine release test with skin test (Phazet), RAST (Phadebas) and bronchial provocation test in children with allergic asthma. The study comprised 68 selected children with a case history of extrinsic allergic asthma to danders (cat and dog) and house-dust mite. Skin prick test, RAST, and histamine release were performed in all children and the bronchial provocation test was used as a reference of "true allergic asthma". A total of 81 allergen bronchial challenges were performed and 44 children experienced 49 positive provocations. In 2.9% (2/68) of the children histamine release could not be performed due to technical difficulties (low histamine release with anti-IgE). Concordances in the range 76-87% were observed with no significant difference between the tests. The highest concordance (87%) was found between histamine release and bronchial provocation test followed by skin prick test vs bronchial provocation (84%) and RAST vs bronchial provocation (80%). The sensitivity and specificity were calculated for each test. All tests showed sensitivities in the range 90-94% and no significant difference between them was observed. The specificity of histamine release, skin prick test, and RAST was 0.78, 0.69, and 0.63, respectively. The specificity of histamine release was better than RAST demonstrated by 95% confidence intervals. In conclusion, it was found that the histamine release test is a convenient diagnostic method and the study indicates a diagnostic value comparable to the common diagnostic methods in clinical allergy.

Animals↗

PubMed1999

[Immunobiology of respiratory allergy].

Respiratory allergies are frequent and the etiological diagnosis is not always easy. Most often, the allergy assessment is vital. For the allergist, this assessment begins with skin tests which must be complemented by measurement of the level of specific IgE in case of doubt. For the generalist or physician who does not have special competence in allergy, an alternative to a specialist consultation at once is a multi-allergen test of detection. This assessment is necessary to set out the etiological diagnosis and facilitate the responsibility of the therapy.

Allergens↗

PubMedDate not supplied

Anaphylaxis caused by cloxacillin: diagnosis with seriated analysis by way of basophil activation test.

We report a case of anaphylaxis caused by cloxacillin in a 13-year-old patient. The basophil activation test, performed 25 days after the anaphylactic reaction, was positive to cloxacillin, amoxicillin, and penicillin G and negative to ibuprofen, tolerated by the patient. The analysis was performed 17 days after the reaction was not conclusive because 74% of the basophil population was activated in basal conditions. The abnormally high activation was similar to that found in an analysis before the reaction, exactly 4 days after finishing a well-tolerated treatment with amoxicillin. This first analysis was available because a patient's sample was taken from the emergency laboratory as a blind control for a study to assess the basophil activation test reliability in diagnosis of hypersensitivity to NSAIDs. The high number of activated basophils in basal conditions after treatment with amoxicillin and before the anaphylactic reaction to cloxacillin probably reflects the beginning of the sensitization. Until now, no cases of hypersensitivity to cloxacillin have been diagnosed by means of the basophil activation test.

Adolescent↗

PubMed1999

Effect of two different types of vacuum cleaners on airborne Fel d 1 levels.

BACKGROUND: Vacuum cleaners may increase the level of airborne allergens by leakage through the cleaners or by disturbance of floor dust by the exhaust air produced. OBJECTIVE: The aim of this study was to evaluate the short-term effect of vacuum cleaning with two different types of cleaners on airborne cat allergen analyzed by a biologic and by an immunochemical test. METHODS: Ten homes with cats were cleaned in random order with a 1-week interval by a traditional canister type vacuum cleaner (T) and a semi-stationary vacuum cleaner (S) that conducts the air to the exterior through a valve in the wall. Airborne particles were collected by air sampling for 2 hours and cat allergen, Fel d 1, was quantified biologically by basophil histamine release test (HR test) and immunochemically by enzyme linked immunosorbent assay (ELISA). RESULTS: Using the S resulted in smaller amounts of airborne cat allergen than the T (mean 2.1 ng/m3 air (range .8 to 12.5) versus 5.2 ng/m3 (1.3 to 13.3), P < .002 measured by ELISA). Results from ELISA and HR test correlated well (r = .91, P < .001). CONCLUSIONS: The use of S with exhaust to the outside of the dwelling gave rise to less airborne low particle size allergen during the cleaning procedure than a T method. The basophil histamine release test could be a valid alternative method to establish allergen content in environmental samples especially in allergen systems with no available monoclonal antibodies.

Air Pollutants↗

PubMed2006

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

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.

Allergens↗

PubMed2006

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.

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.

Adolescent↗

PubMed1997

[Immuno-biological diagnosis of food allergy].

Food allergy is becoming more frequent, with 6% of asthmatics reporting an isolated food allergy, and 5 to 6% of atopic dermatitis patients also have either a single or multiple true food allergy. There is value in immuno-biological diagnosis by: Measurement of total serum IgE. Measurement of mono-allergen-specific IgE, following a measurement by a multi-allergen of the Trophatope type. A study of elimination of foods for 2 or 3 months followed by their re-introduction. Oral provocation tests in a hospital environment under clinical control and subsequent measurement of the mediators:-Plasma histamine, tryptase, and urinary methylhistamine to give proof of responsibility of the food allergen. Nowadays, it is perfectly possible to include in diagnosis the new technologies of the test of activation of basophils/or lymphocytes by means of flow cytometry.

Adolescent↗

PubMed1997

[Anaphylactic or anaphylactoid reactions to paracetamol. Report of 3 cases].

Anaphylaxis to paracetamol exists and perhaps nowadays diagnosis is not confined to clinical evidence of: The repeatability of clinical symptoms. Skin tests. Provocation tests. But also by immuno-biological evidence. Application of the test of Activation of basophils by Flow Cytometry and measurement of leukotriene C4 have been nowadays largely validated and constitute irrefutable proof of the responsibility of the drug as the origin of the clinical symptoms.

Acetaminophen↗

PubMed2003

[Basophil activation test in the diagnosis of allergy to medicines].

In this paper we study the reliability of the basophil activation test (BAT) in the "in-vitro" diagnosis of allergy to betalactams and to metamizol, and the sensitivity and specificity of the technique are analyzed. To this end, we studied 58 patients allergic to betalactam antibiotics with a positive cutaneous test facing any derivative of penicillin and 30 healthy controls who tolerated betalactams, and 26 patients allergic to metamizol with an immediate reaction and 30 healthy controls who tolerated the medicine. Sensitivity to BAT in allergy to betalactams was 52.8%, and specificity was 92.6%. For metamizol, sensitivity was 42.3% and specificity was 100%. The positive predictive value of BAT in allergy to betalactams was 18.9% and the negative predictive value was 98.4%. For metamizol, the positive predictive value of the technique was 100% and the negative predictive value was 99.4%. The joint use of BAT and CAP (specific IgE) makes it possible to diagnose some 65% of patients allergic to betalactams. The combined use of cutaneous tests and BAT in allergy to metamizol detects 70% of the cases. BAT is a useful, non-invasive technique in the "in-vitro" diagnosis of allergy to betalactams and metamizol.

Basophil Degranulation Test↗

PubMedDate not supplied

[Drug allergy assessed by modified Shelley's test. A case series].

BACKGROUND: The frequency of allergy to drugs is 5-10% in the general population, The Shelley's test is a method in vitro for its diagnosis and it is considered positive when it is equal o greater than 20%. OBJECTIVE: To show the allergy to drugs frequency evaluated by the Shelley's Modified Test. MATERIAL AND METHODS: We included 165 patients with clinical diagnosis of allergy to drugs to those who the test of Shelley's was applied modified test for one or several medicines. The study was made in the period of January 1st to December 3st 2002. RESULTS: 88% (145 patients) were positive, and 12% (20 patients) were negative in correlation with clinical findings. The female was more determinant (2:1 relation), with an average age of 45 years. The drugs that caused reactions of allergy in were the antibiotics (sulfas, penicillin, quinolones), analgesic (ASA, naproxen, diclofenac), antihypertensive (enalapril, captopril, verapamil) and others with 95, 170, 48 and 97 cases respectively. The most frequent clinical features were urticaria, angioedema, anaphylactic shock and erythema. CONCLUSION: The Shelley's test is useful to support the diagnosis of allergy to drugs, although does not exclude it. The antibiotics intake is the most common cause and the most common clinical manifestation was urticaria.

Adolescent↗

PubMed1994

Antiallergic activity of tylogenin, a novel steroidal compound from Tylophora sylvatica.

Tylogenin, a steroidal aglycone generated by acid hydrolysis from two seasonal glycosides occurring in Tylophora sylvatica, inhibits IgE-induced basophil mediator release for allergic reactions. In the rabbit basophil-dependent serotonin release (BDSR) assay system, the inhibitory activity of tylogenin (geom mean IC50 = 39 microM) was significantly (P < 0.05) greater than that of its parent glycosides, tylophoroside (geom mean IC50 263 microM) and acetyltylophoroside (geom mean IC50 331 microM), and that of dexamethasone (geom mean IC50 = 912 microM). The activity of tylogenin was found to increase with the incubation time. In the human leukocyte-dependent histamine release (LDHR) test model, the glycosides had only a minimal activity. In contrast, tylogenin, with a geom mean IC50 = 49 microM, exerted a significantly (P < 0.05) greater potency than dexamethasone (IC50 = 257 microM). These results suggest that tylogenin could represent a new class of antiallergic agents.

4-Butyrolactone↗

PubMed1999

[Level of histamine in supernatants from the basophil activation test: applications to hymenoptera allergy and drug allergy--preliminary study].

Histamine Release technic consists in calculating histamine liberated by blood cells in touch with an allergen. To this day, this method is only used in Hymenoptera venom allergy diagnosis. The principle of this study is to measure histamine released by activated basophils in surnageons of Basophil Activation Test (BAT) for different allergens: Hymenoptera venoms: Bee, White Faced Hornet, Vespula Wasp. Drugs: Cefaperos, Clamoxyl, Alfatil, Rapifen, Diprivan, Nesdonal, Mivacron. A threshold of positivity (amplification factor in comparison with the control) is determined for these two classes of allergens: 45 for Hymenoptera venoms and 9 for drugs. These results, compared to the other diagnosis technics (Histamine Release, Basophil Activation Test, Prick Tests) discloses very high correlation rates in each case. This method seems to be a reliable method for Hymenoptera venom allergy diagnosis and for drugs allergy diagnosis too. However, this study is based on a few number of patients, so a significant statistic conclusion can't be expressed but it opens an interesting way of research.

Basophil Degranulation Test↗

PubMed2005

Drug hypersensitivities: which room for biological tests?

Drug allergic reactions frequently represent a serious diagnostic problem. In this paper we summarise the most relevant data published in recent years on the diagnostic reliability of the in vitro techniques in drug allergy diagnosis. The lymphocyte transformation test (LTT) offers a sensitivity of 58% in the diagnosis of late allergic reactions to betalactams and 64.5% in the immediate allergic reactions. The basophil activation test and the antigen-specific sulphidoleukotriene determination have an acceptable diagnostic reliability in muscle relaxant drug-induced reactions and in betalactam allergy. BAT sensitivity in betalactam allergy was 50.7% and its specificity 93.3%, whereas CAP had a sensitivity of 36.7% and a specificity of 83.3%, and CAST, a sensitivity of 47.7% and a specificity of 83.3%. For NSAID hypersensitivity, BAT sensitivity was 63.3% and specificity 93.3%, CAST sensitivity was 38.3% and specificity 76.6%. BAT sensitivity in metamizol allergy was 42.3% and the specificity 100% and CAP was negative in all the 17 cases in which it was performed. The joint use of BAT and CAP (specific IgE) allows diagnosis of 65.2% of the betalactam allergic patients with a specificity of 83.3%. The combined use of CAST and BAT in metamizol allergy detects 76% of the cases and 76.9% when associating the skin tests. In NSAID hypersensitivity, the joint use of BAT and CAST does not increase the diagnostic reliability of BAT alone. BAT is a non-invasive useful technique in the in vitro diagnosis of betalactam and metamizol allergy, and NSAID hypersensitivity.

Anti-Bacterial Agents↗

PubMed2003

Anaphylactic reaction after artificial insemination.

BACKGROUND: Bovine seroalbumin is known as an allergen for human beings, but reactions to it in an artificial insemination procedure are much rarer. We report a case of anaphylaxis after intrauterine insemination (IUI) in which sensitization to bovine serum albumin (BSA) is demonstrated. OBJECTIVE: Report the allergy evaluation performed in a patient who suffered a severe reaction immediately after an IUI procedure. METHODS: A 33-year-old woman was referred because of an anaphylactic reaction after a second trial of IUI. She developed pruritus, abdominal pain, nausea and vomiting, bronchospasm, and generalized urticaria. She had an atopic medical history of pollen allergy and sensitization to cat epithelium. She had never had trouble with minor surgery and she usually uses latex material. She had never received heterologous sera before. Her husband's semen for the IUI was processed in a standard fluid medium called upgraded INRA B 2 (Laboratoires CCD, Paris, France), which contains amino acids, lipids, vitamins, BSA, penicillin, and streptomycin in addition to inorganic salts. RESULTS: Skin prick tests with the medium and BSA 10 mg/mL were positive. In vitro studies demonstrated an immunoglobulin E binding protein of 60 to 65 kDa and mast cells and basophil activation (CD63 expression) against BSA contained in the medium. Cutaneous and challenge tests with penicillin and streptomycin were negative. CONCLUSIONS: We consider the BSA in the semen culture medium to be the factor which triggered the anaphylactic reaction. This case supports the authors who state that media free from heterologous proteins should be used for human application, especially on atopic patients, to avoid sensitization.

Adult↗

PubMed2006

Diagnosis of neuromuscular blocking agent hypersensitivity reactions using cytofluorimetric analysis of basophils.

BACKGROUND: Immunoglobulin E (IgE)-mediated hypersensitivity reactions to neuromuscular blocking agents (NMBA) are common and life threatening. Basophil activation based upon the expression of CD63 in the presence of specific allergens was found to be of importance for the diagnosis of IgE-mediated hypersensibility. METHODS: The Basotest was evaluated for the diagnosis of NMBA in 47 patients with proven NMBA anaphylaxis, 40 atopic subjects nonallergic to NMBA and five healthy volunteers. Diagnosis of NMBA was made according to international standards on clinical history, skin tests and provocation tests when needed. RESULTS: In the NMBA allergic patients, sensitivity of Basotest was 36.1%, but it increased to 85.7% for reactions which occurred within the last 3 years. The specificity was 93.3%. CONCLUSION: Basotest may be useful for the diagnosis of NMBA allergy in patients with a suspicion of recent IgE-mediated hypersensitivity reaction to NMBA.

Adult↗

PubMed1993

Lack of role for mononuclear cell-derived histamine releasing factors in occupational asthma due to western red cedar.

Occupational asthma due to Western Red Cedar (WRCA) is attributed to sensitization to plicatic acid (PA), but does not appear to be dependent on PA-specific IgE antibodies. Exposure to PA induces histamine release in vivo and in vitro, so if IgE is not important, other mechanisms of histamine release must presumably operate in WRCA. To explore the possible role of histamine-releasing factors in WRCA, peripheral blood mononuclear cells were obtained and cultured with PA, PA-albumin conjugate plicatic acid-human serum albumin (PA-HSA), grass pollen or Concanavalin A using a standard histamine releasing factor (HRF) generation protocol. Supernatants were dialysed to remove endogenous histamine and then assayed for histamine releasing activity using human basophils as targets and a Con A-induced bulk supernatant as an internal HRF standard. In contrast to some previous reports, spontaneous HRF release from the peripheral blood mononuclear cells (PBMC) of WRCA patients (n=9) and atopic asthmatic subjects (n=5) was not elevated compared with the non-asthmatic controls (n=11; five atopic and six non-atopic). Both PA and PA-HSA induced the production of small amounts of HRF by PBMC of WRCA patients, but a similar degree of HRF generation was also observed in PBMC from the atopic asthmatic, atopic nonasthmatic, and non-atopic subjects. In contrast, grass pollen induced the production of HRF by PBMC from the subjects with positive skin tests to grass pollen but not by PBMC of non-atopic subjects, confirming that our methods and assay were capable of detecting antigen-specific HRF production. Since neither PA nor PA-HSA induced significantly elevated HRF production from PBMC of WRCA patients, it seems unlikely that PA-induced HRFs play a substantial role in the pathogenesis of WRCA.

Adult↗

PubMedDate not supplied

Eosinophilic esophagitis.

BACKGROUND: Esophagitis is an increasingly diagnosed disease. Patients with gastroesophagic reflux, dysphagia, vomiting or abdominal pain, with a torpid response to the treatment, could be suffering from it. MATERIAL AND METHODS: A 37 year-old male patient with background of gastroesophagic reflux and dysphagia for solids since 2002, self-limited diarrhea episodes and intolerance to alcoholic drinks due to epigastric pain. Skin prick tests, specific IgE, histamine release test and basophil activation test were carried out. RESULTS: Skin prick test to the usual allergens with negative result; prick-prick tests to egg white and yolk, milk and apple with positive result to egg white; total serum IgE within normal levels, specific IgE to egg white with positive result; histamine release test (HRT) and basophil activation test (BAT) with positive result to egg white and yolk. CONCLUSION: The patient was diagnosed eosinophilic esophagitis. The commercial food extracts have a great variability in their allergenic composition, which could result in false negative results in the prick test. Prick-prick with the natural food is a more sensitive technique than prick in the diagnosis of food allergy. There are other useful in vitro techniques, apart from specific IgE, in the diagnosis of food allergy. In our case, an exclusion diet of the involved food was more effective than other treatments for remission of the symptoms.

Adult↗