Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor FcɛRI

Sabban, Sari; Ye, Hongtu; Helm, Birgit A.

doi:10.1016/j.vetimm.2013.01.008

Cited by 12 publications

(18 citation statements)

References 32 publications

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“…Graph B shows the release assay preformed on RBL-2H3.1 parental cells which not express equine FcεRIα and those expressing equine FcεRIα, using equine IgE anti NIP-HSA for cell sensitization. The equine IgE binds to the equine receptor, but not the endogenous rat receptor, to result in mediator release, this is confirmed since the parental cells did not release mediator mediators, while the cells expressing the equine receptor support equine IgE-mediated, antigen induced, mediator release 2,3 .…”

Section: Representative Resultsmentioning

confidence: 71%

“…A1 and A7 represent the location of the wells in the 96 well plate. Plot a graph of percentage of β-hexosaminidase release (which corresponds to total mediator release) against antigen concentration 2,3 .…”

Section: ) Release Assaymentioning

confidence: 99%

“…This assay was modified from previous assays used to study human and canine allergic responses 4,5 . We have also shown that this type of assay system has multiple applications for the development of diagnostic tools and the safety assessment of potential therapeutic intervention strategies in allergic disease 6,2,3 . …”

mentioning

confidence: 98%

“…The gene product of the transfected equine α-chain formed a functional receptor complex with the endogenous rat β-and γ-chains 1 . The resultant assay system facilitated an assessment of the quantity of mediator secreted from equine FcεRIα transfected RBL-2H3.1 cells following sensitization with equine IgE and antigenic challenge using β-hexosaminidase release as a readout 2,3 . Mediator release peaked at 36.68% ± 4.88% at 100 ng ml -1 of antigen.…”

mentioning

confidence: 99%

“…In this article, we now present such an assay system applicable to the study of diagnostic and therapeutic strategies relevant to the equine system, where β-hexosaminidase release, as an indicator of cell mediator degranulation, was assessed on RBL-2H3.1 cells expressing equine FcεRIα. This protocol is based on previous publications 25,4,5,2,3 describing the engineering of RBL cells transfected with the gene encoding the IgE binding domain of the high-affinity receptor for IgE from different species. The protocol explains how to perform a β-hexosaminidase release assay, the results of which are presented as the mean ± standard deviation of triplicate experiments.…”

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confidence: 99%

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Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor FcεRI

Sabban

Helm

2014

JoVE

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Citation: Sabban, S., Ye, H., Helm, B. Development of an in vitro model system for studying the interaction of Equus caballus IgE with its highaffinity receptor FcεRI. J. Vis. Exp. (93), e52222, doi:10.3791/52222 (2014). AbstractThe interaction of IgE with its high-affinity Fc receptor (FcεRI) followed by an antigenic challenge is the principal pathway in IgE mediated allergic reactions. As a consequence of the high affinity binding between IgE and FcεRI, along with the continuous production of IgE by B cells, allergies usually persist throughout life, with currently no permanent cure available. Horses, especially race horses, which are commonly inbred, are a species of mammals that are very prone to the development of hypersensitivity responses, which can seriously affect their performance. Physiological responses to allergic sensitization in horses mirror that observed in humans and dogs. In this paper we describe the development of an in situ assay system for the quantitative assessment of the release of mediators of the allergic response pertaining to the equine system. To this end, the gene encoding equine FcεRIα was transfected into and expressed onto the surface of parental Rat Basophil Leukemia (RBL-2H3.1) cells. The gene product of the transfected equine α-chain formed a functional receptor complex with the endogenous rat β-and γ-chains 1 . The resultant assay system facilitated an assessment of the quantity of mediator secreted from equine FcεRIα transfected RBL-2H3.1 cells following sensitization with equine IgE and antigenic challenge using β-hexosaminidase release as a readout 2,3 . Mediator release peaked at 36.68% ± 4.88% at 100 ng ml -1 of antigen. This assay was modified from previous assays used to study human and canine allergic responses 4,5 . We have also shown that this type of assay system has multiple applications for the development of diagnostic tools and the safety assessment of potential therapeutic intervention strategies in allergic disease 6,2,3 . Video LinkThe video component of this article can be found at

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Section: Representative Resultsmentioning

confidence: 71%

Section: ) Release Assaymentioning

confidence: 99%

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Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor FcεRI

Sabban

Helm

2014

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Are humanized IgE reporter systems potential game changers in serological diagnosis of human parasitic infection?

et al. 2021

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Immunoglobulin E (IgE) is thought to have evolved to protect mammalian hosts against parasitic infections or toxins and plays a central role in the pathogenesis, diagnosis, and therapy of IgE-mediated allergy. Despite the prominence of IgE responses in most parasitic infections, and in stark contrast to its use in the diagnosis of allergy, this isotype is almost completely unexploited for parasite diagnosis. Here, we discuss the perceived or real limitations of IgE-based diagnosis in parasitology and suggest that the recent creation of a new generation of very sensitive cellular IgE-based reporters may represent a powerful new diagnostic platform, but needs to be based on a very careful choice of diagnostic allergens.

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Detailed Analysis of Kinetic Binding Traces with Distributions of Surface Sites

Zhao¹,

Schuck²

2017

Handbook of Surface Plasmon Resonance

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In kinetic surface binding experiments, deviations from the ideal exponential binding traces are commonly observed. Often, these can be attributed to the intrinsic heterogeneity of the binding properties of the immobilized molecules. Surface binding traces can be fitted with a mathematical model consisting of a heterogeneous distribution of surface sites with quasi-continuous affinity constants and kinetic rate constants. It is implemented in the software EVILFIT both for reaction-controlled binding and, if required, with approximate corrections in the onset of mass transport limitation. Statistical tools prevent over-interpretation and highlight the information content of the data. Examples illustrate impact of the choice of surface chemistry on the surface sites, and also factors such as immobilization density and time after immobilization.

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Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor FcɛRI

Cited by 12 publications

References 32 publications

Development of an <em>in vitro</em> model system for studying the interaction of <em>Equus</em> <em>caballus</em> IgE with its high-affinity receptor FcεRI

Development of an <em>in vitro</em> model system for studying the interaction of <em>Equus</em> <em>caballus</em> IgE with its high-affinity receptor FcεRI

Are humanized IgE reporter systems potential game changers in serological diagnosis of human parasitic infection?

Detailed Analysis of Kinetic Binding Traces with Distributions of Surface Sites

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Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor FcɛRI

Cited by 12 publications

References 32 publications

Development of an <em>in vitro</em> model system for studying the interaction of <em>Equus</em> <em>caballus</em> IgE with its high-affinity receptor Fc&#949;RI

Development of an <em>in vitro</em> model system for studying the interaction of <em>Equus</em> <em>caballus</em> IgE with its high-affinity receptor Fc&#949;RI

Are humanized IgE reporter systems potential game changers in serological diagnosis of human parasitic infection?

Detailed Analysis of Kinetic Binding Traces with Distributions of Surface Sites

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Development of an <em>in vitro</em> model system for studying the interaction of <em>Equus</em> <em>caballus</em> IgE with its high-affinity receptor FcεRI

Development of an <em>in vitro</em> model system for studying the interaction of <em>Equus</em> <em>caballus</em> IgE with its high-affinity receptor FcεRI