Mediator release assays based on human or murine IgE in allergen standardization

Kaul, Susanne; Hoffmann, Andreas; Lüttkopf, D.; Kastner, B.; Höltz, G.; Vieths, S.

doi:10.1016/s0091-6749(03)80859-x

Cited by 3 publications

(5 citation statements)

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“…The correlation of mediator release with data from immunochemical analysis and clinical observation is very good, provided that the sera used in the biological test system are evaluated and carefully chosen (Hoffmann et al, 2003;Kaul et al, 2006). This is no problem for more prevalent allergies, since enough patients can be recruited for blood donation.…”

Section: Discussionmentioning

confidence: 99%

“…Despite this drawback, the mediator release assay gives consistent results with coefficients of variation between 10-15 % and a sensitivity that can detect low amounts of allergenic proteins (10 -9 -10 -11 g) and in some cases even in the femtogram-range (Hoffmann et al, 1995). The comparison of three cell-based assays, namely mediator release from RBL-2H3 or transfected RBL-30/25 cells and stripped human basophils, respectively, resulted in almost identical data (Hoffmann et al, 2003;Kaul et al, 2006). Since the method using stripped basophils requires freshly prepared human cells, its application in routine analysis is restricted for practical and ethical reasons.…”

Section: Discussionmentioning

confidence: 99%

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Development of a Biological Assay to Determine the Allergenic Potential of Foods

2006

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Abbreviations: ELISA = enzyme-linked immunosorbent assay; PCR = polymerase chain reaction; SDS-PAGE = sodium dodecyl sulfatepolyacrylamide gel electrophoresis; FcεRI = high affinity receptor for IgE; RBL = rat basophilic leukemia; PBS = phosphate-buffered saline; TBS = tris-buffered saline.Abstract: Food allergies represent a risk for many people in industrialized countries. Unrecognizable allergenic proteins of foodstuffs may be present as ingredients that are not labeled or as unknown cross-contamination. Such hidden allergens can cause severe reactions in allergics, even at minute quantities, sometimes with fatal outcome. For the verification of the presence of allergenic food constituents, analytical methods such as ELISA and PCR have been developed. However, these tests cannot measure allergenic potential. For this reason, a test system that measures the biological activity of allergens has been developed. It is based on the cellular mechanisms of the type I allergy. Rat basophilic leukemia cells (RBL-2H3) were transfected with the genes of the human high affinity receptor for IgE. The resulting cell line expressed the human receptor α-chain and could bind allergenspecific IgE from allergic subjects, in contrast to the parent cell line. After cross-linking of receptor-bound, allergen-specific human IgE by allergens, the cells released measurable inflammatory mediators. These cells were used for the analysis of a variety of allergen extracts, including extracts prepared from foods containing allergenic hazelnut and peanut. The comparative validation with existing ELISA and PCR for hazelnut and peanut demonstrated similar sensitivity and specificity. The established cell line will be a novel tool in the detection of allergens in complex mixtures, especially to address the issue of their allergenic potential, which cannot be accomplished by classical analytical methods. This will add valuable information about the allergenic potential of food constituents to the risk assessment of foods.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Development of a Biological Assay to Determine the Allergenic Potential of Foods

2006

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“…In this context, it should be noted that effector cell assays such as the basophil histamine release test [24,25] utilize the same mechanism as skin tests and thus would reflect the biological activity of allergens more closely. Such tests, however, are technically demanding and much more difficult to perform, and, in general, the precision of biological test systems is lower than that of antibody binding tests.…”

Section: Immunological and Biochemical Methods In Allergen Standardizmentioning

confidence: 99%

“…In the field of measuring the biological potency of allergens and allergen extracts, cellular assays have been developed that mimic the effector mechanism of type I allergies, using either stripped and resensitized basophils [24,25], a rat basophilic leukaemia cell line (RBL-2H3) or RBL cells transfected with the genes of the human high-affinity IgE receptor [25,43 ]. These cell lines could be passively sensitized with murine (RBL-2H3) or human IgE (transfected RBL and stripped basophils), obtained from immunized mice or the serum of allergic patients, respectively.…”

Section: Recent Findings In the Field Of Allergen Standardizationmentioning

confidence: 99%

“…These cell lines could be passively sensitized with murine (RBL-2H3) or human IgE (transfected RBL and stripped basophils), obtained from immunized mice or the serum of allergic patients, respectively. In the study by Kaul et al [25], different types of mediator release assays were used to analyse the potency of extracts from birch and timothy pollen, house dust mite, cat dander and peanut, and the results were compared with those of a validated IgE inhibition assay. As a good correlation with inhibition data, a very high analytical sensitivity and a relatively high precision of the biological test were obtained (coefficient of variation below 15%), the authors concluded that mediator release tests may be suitable to determine the biological activities of allergens or allergen extracts.…”

Section: Recent Findings In the Field Of Allergen Standardizationmentioning

confidence: 99%

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Standardization of allergen extracts for immunotherapy: where do we stand?

Becker

Vogel

Vieths

2006

Current Opinion in Allergy &Amp; Clinical Immunology

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Several new tests have been developed to make more major and minor allergens from various allergen sources accessible to allergen standardization programmes such as the CREATE project. It is expected that assays to determine the majority of all clinically relevant major allergens from aeroallergen sources will be available in the near future. Standardized and validated mediator release assays may be a complementary tool for evaluating the biological potency of reference allergens and for correlating allergen concentrations to biological potency.

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