Measurement of basophil‐activating capacity of grass pollen allergens, allergoids and hypoallergenic recombinant derivatives by flow cytometry using anti‐CD203c

Kahlert, Helga; Cromwell, Oliver; Fiebig, Helmut

doi:10.1046/j.1365-2222.2003.01756.x

Cited by 48 publications

(54 citation statements)

References 21 publications

(28 reference statements)

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“…The most prominent markers are the tetraspanin CD63 [46] and the ecto-enzyme CD203c [47][48][49]. Both markers are suitable to analyze allergen-induced basophil activation and are routinely used in allergy diagnosis [19][20][21][22][23][24][25][26][27][28][29][30]. In the search for additional markers, we analyzed a panel of workshop antibodies for differential reactivity with resting and activated basophils.…”

Section: Discussionmentioning

confidence: 99%

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Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation

Hennersdorf¹,

et al. 2005

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Using two-colour flow cytometry >200 antibodies submitted to the 8 th International Workshop of Human Leukocyte Differentiation Antigens (HLDA8) have been analyzed for their reactivity with resting and activated CD203c + basophils. Four antibodies either non-reactive or weakly reactive with resting basophils exhibited an increased reactivity with basophils activated by anti-IgE-mediated cross-linking of the high affinity IgE receptor (FcεRI). These include antibodies against CD164 (WS-80160, clone N6B6 and WS-80162, clone 67D2), as well as two reagents with previously unknown specificities that were identified as CD13 (WS-80274, clone A8) and CD107a (WS-80280, clone E63-880). The activation patterns followed either the "CD203c-like" or "CD63-like" activation profile. The CD203c profile is characterized by a rapid and significant upregulation (of CD13, CD164, and CD203c), reaching maximum levels after 5-15 min of stimulation. The phosphoinositide-3-kinase (PI3K)-specific inhibitor wortmannin inhibited the upregulation of these markers whereas 12-O-tetradecanoyl-phorbol-13-acetate (TPA) induced a rapid and FcεRI-independent upregulation within 1-2 min. In the CD63 profile, maximum upregulation (of CD63 and CD107a) was detected only after 20-40 min, and upregulation by TPA reached maximum levels after 60 min. In summary, our data identify CD13, CD107a, and CD164 as novel basophil-activation antigens. Based on time kinetics of upregulation, we hypothesize that molecules of the "CD203c group" and the "CD63 group" are linked to two different mechanisms of basophil activation.

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Section: Discussionmentioning

confidence: 99%

“…In addition, activation markers like the granuleassociated molecule CD63 and the ecto-enzyme CD203c are upregulated to the plasma membrane in response to FcεRI cross linkage [19][20][21][22][23][24][25][26][27][28][29][30]. These markers are now routinely used for flow cytometry-based basophil activation tests.…”

Section: Introductionmentioning

confidence: 99%

Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation

Hennersdorf¹,

et al. 2005

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“…They are usually generated by incubating the native extracts in the presence of cross-linking agents like formaldehyde, glutaraldehyde or other substances reacting with ε-amino acids [13], which results in the formation of more or less large protein complexes. These complexes show a fundamentally reduced capacity to bind IgE [14, 15]depending on the size of the formed complexes, thereby minimizing the release of anaphylaxis-inducing mediators. The higher the molecular weight, the lower the risk of side effects [16].…”

Section: Introductionmentioning

confidence: 99%

Characterization of Modified Allergen Extracts by in vitro β-Hexosaminidase Release from Rat Basophils

Gehlhar

Peters

Brockmann

et al. 2005

Int Arch Allergy Immunol

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Background: To date, there is no well-established test available that can be used to measure functional properties of modified allergens (allergoids). Due to the cross-linking process, the IgE-binding capacity of the allergens, normally necessary for their characterization, is lost. The aim of this study was to test whether the rat basophilic leukaemia (RBL) cell assay (β-hexosaminidase release by rat basophils upon allergen stimulation) can be adopted to characterize allergoids and to evaluate the assay for testing allergoids and native allergens as well. Methods: Mice were immunized with native and modified Phleumpratense extracts in the presence of alum. Their sera were used to sensitize RBL-2H3 cells and measure basophil stimulation induced by different allergen extracts in the presence or absence of various additives. Results: Sera containing specific IgE against both extract formulations were obtained. Native as well as modified extracts induced dose-dependent β-hexosaminidase release from RBL cells. Both extracts were used to evaluate the characteristics of the assay, which showed high precision. Storage conditions were chosen to enhance extract degradation, which could be read directly from the altered stimulatory capacity of the extracts. Additives turned out to have diverse effects on the assay, whereas phenol had no measurable effect, alum had an inhibitory effect and glycerol elevated basophil activation. Conclusions: For the first time, a reliable, precise in vitro assay is available that is able to directly measure the properties of modified allergen extracts after their production process. The test is well evaluated and its advantages and limitations are discussed in this report.

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“…In the subsequent research performed to optimize this technique, other cellular markers such as the marker of activation of basophils CD203c were found. Since the mid-1990s, BAT based on CD63 and CD203c has proven reliable for the diagnosis of IgE-mediated allergies (Buhring et al, 2004;Hauswirth et al, 2002;Kahlert et al, 2003;Mayorga et al, 2010;Ocmant et al, 2009). As compared with routine diagnostic tests specific IgE or SPT, BAT was less sensitive, particularly when including nonresponders, but was more specific, achieving at least 95% specificity (Ocmant et al, 2009).…”

Section: Human Basophil Modelmentioning

confidence: 99%

Use of a rat basophil leukemia (RBL) cell-based immunological assay for allergen identification, clinical diagnosis of allergy, and identification of anti-allergy agents for use in immunotherapy

Sun

Zhou

Zhou³

et al. 2014

Journal of Immunotoxicology

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(2015) Use of a rat basophil leukemia (RBL) cell-based immunological assay for allergen identification, clinical diagnosis of allergy, and identification of anti-allergy agents for use in immunotherapy, Journal of Immunotoxicology, 12:2, 199-205, DOI: 10.3109/1547691X.2014 AbstractFood allergy is an important public health problem that affects an estimated 8% of young children and 2% of adults. With an increasing interest in genetically-engineered foods, there is a growing need for development of sensitive and specific tests to evaluate potential allergenicity of foods and novel proteins as well as to determine allergic responses to ensure consumer safety. This review covers progress made in the field of development of cell models, specifically that involving a rat basophil leukemia (RBL) cell-based immunoassay, for use in allergen identification, diagnosis, and immunotherapy. The RBL assay has been extensively employed for determining biologically relevant cross-reactivities of food proteins, assessing the effect of processing on the allergenicity of food proteins, diagnosing allergic responses to whole-food products, and identifying anti-allergy food compounds. From the review of the literature, one might conclude the RBL cell-based assay is a better test system when compared to wild-type mast cell and basophil model systems for use in allergen identification, diagnosis, and analyses of potential immunotherapeutics. However, it is important to emphasize that this assay will only be able to identify those allergens to which the human has already been exposed, but will not identify a truly novel allergen, i.e. one that has never been encountered as in its preferred (humanized) configuration.

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Measurement of basophil‐activating capacity of grass pollen allergens, allergoids and hypoallergenic recombinant derivatives by flow cytometry using anti‐CD203c

Cited by 48 publications

References 21 publications

Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation

Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation

Characterization of Modified Allergen Extracts by in vitro β-Hexosaminidase Release from Rat Basophils

Use of a rat basophil leukemia (RBL) cell-based immunological assay for allergen identification, clinical diagnosis of allergy, and identification of anti-allergy agents for use in immunotherapy

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