Identification of amino acid residues involved in the interaction of canine IgE with canine and human FcɛRIα

Ye, Hongtu; Housden, Jonathan E.M.; Hunter, Malcolm; Sabban, Sari; Helm, Birgit A.

doi:10.1016/j.molimm.2013.08.013

Cited by 6 publications

(6 citation statements)

References 26 publications

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“…Two heavy chains of one IgE molecule have different amino acid sequences involved in contacts with the receptor [26]. Subdomains reported to contact the receptor FceRI include a linker connecting the IgE domains Ce2 and Ce3 (Ce2-Ce3 linker, residues 334-336), and the Ce3 loops BC (residues 362-365), DE (393-396), and FG (424-427) [28,30,31]. The FceRI molecule has two cavities, namely P426 and Y131 pockets [26].…”

Section: Discussionmentioning

confidence: 99%

SplitCore Technology Allows Efficient Production of Virus-Like Particles Presenting a Receptor-Contacting Epitope of Human IgE

et al. 2015

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Section: Discussionmentioning

confidence: 99%

SplitCore Technology Allows Efficient Production of Virus-Like Particles Presenting a Receptor-Contacting Epitope of Human IgE

et al. 2015

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“…[84][85][86][87] Such humanized RBL cells were then used to test the capacity of purified peanut allergens to induce degranulation [88][89][90] or food allergens. 91 Since then, the RBL cells have also been transfected to express equine 92 and canine 93 high affinity IgE receptor α chains, extending their use to other mammalian species.…”

Section: Rbl-sx-38; Rbl-48; Rbl30/25: the Human Fcεriα Expressing Rmentioning

confidence: 99%

RBL cells as models for in vitro studies of mast cells and basophils

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Barwary

et al. 2018

Immunological Reviews

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Since their establishment in 1981, RBL-2H3 cells have been widely used as a mast cell (MC) model. Their ability to be easily grown in culture in large amounts, their responsiveness to FcεRI-mediated triggers and the fact that they can be genetically manipulated, have provided advantages over primary MCs, in particular for molecular studies relying on genetic screening. Furthermore, the ability to generate clones that stably express proteins of interest, for example, a human receptor, have marked the RBL cells as an attractive MC model for drug screening. Indeed, 3 RBL reporter cell lines (RS-ATL8, NFAT-DsRed, and NPY-mRFP) have been generated providing useful models for drug and allergen screening. Similarly, RBL cells stably expressing the human MrgprX2 receptor provide a unique paradigm for analyzing ligand interactions and signaling pathways of the unique human receptor. Finally, transient co-transfections of RBL cells allow functional genomic analyses of MC secretion by combining library screening with simultaneous expression of a reporter for exocytosis. RBL cells thus comprise powerful tools for the study of intracellular membrane trafficking and exocytosis and the detection of allergens, vaccine safety studies and diagnosis of allergic sensitization. Their recent uses as an investigative tool are reviewed here.

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“…Therefore, in order to assess human sensitization, several groups have created stably transfected humanized RBL cell lines, such as RBL SX-38 [5], RBL 48 [6], RBL 703/21 (which was derived from RBL 30/25 [7]), or RBL hEIa-2B121 [8]. Similarly, RBL-2H3 cell lines have been transfected with non-human receptors allowing allergen-specific IgE measurements in canine [9] and equine [10] blood samples.…”

Section: Introductionmentioning

confidence: 99%

Characterization of human FcεRIα chain expression and gene copy number in humanized rat basophilic leukaemia (RBL) reporter cell lines

et al. 2019

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Several laboratories have created rat basophil leukemia (RBL) cell lines stably transfected with the human high affinity IgE receptor (FcεRI H ). More recently, humanized RBL cell lines saw the introduction of reporter genes such as luciferase (RS-ATL8) and DsRed (RBL NFAT-DsRed). These reporters are more sensitive than their parental non-reporter humanized RBL cell lines. However, no studies so far have addressed the levels of FcεRI H surface expression on humanized RBL cell lines. This is a critical parameter, as it determines the ability of these cells to be efficiently sensitized with human IgE, hence it should affect the sensitivity of the cell assay–a critical parameter for any diagnostic application. Our purpose was to assess and compare the levels of expression of the transfected FcεRI H chain in humanized RBL cell lines. We compared surface levels of FcεRIα H by flow cytometry, using a fluorescently labelled monoclonal antibody (CRA-1/AER-37) and determined receptor numbers using calibration microspheres. FcεRIα H copy numbers were assessed by qPCR, and the sequence verified. Transfection with FcεRIγ H cDNA was assessed for its ability to increase FcεRIα H expression in the NFAT-DsRed reporter. While both SX-38 and RS-ATL8 expressed about 500.000 receptors/cell, RBL 703–21 and NFAT-DsRed had approximately 10- to 30-fold lower FcεRIα H expression, respectively. This was neither related to FcεRI H gene copy numbers, nor to differences in steady state mRNA levels, as determined by qPCR and RT-qPCR, respectively. Instead, FcεRIα H surface expression appeared to correlate with the co-expression of FcεRIγ H . Stable transfection of NFAT-DsRed cells with pBJ1 neo-huFcεRI gamma, which constitutively expresses FcεRIγ H , increased FcεRIα H chain expression levels. Levels of FcεRIα H surface expression vary greatly between humanized RBL reporter cell lines. This difference will affect the sensitivity of the reporter system when used for diagnostic purposes.

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Identification of amino acid residues involved in the interaction of canine IgE with canine and human FcɛRIα

Cited by 6 publications

References 26 publications

SplitCore Technology Allows Efficient Production of Virus-Like Particles Presenting a Receptor-Contacting Epitope of Human IgE

SplitCore Technology Allows Efficient Production of Virus-Like Particles Presenting a Receptor-Contacting Epitope of Human IgE

RBL cells as models for in vitro studies of mast cells and basophils

Characterization of human FcεRIα chain expression and gene copy number in humanized rat basophilic leukaemia (RBL) reporter cell lines

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