MRGPRX2 and Immediate Drug Hypersensitivity: Insights From Cultured Human Mast Cells

Elst, Jessy; Sabato, Vito; Faber, Margaretha A.; Bridts, Chris H.; Mertens, Christel; Houdt, Michel van; Gasse, Athina L. Van; Hagendorens, Margo M.; Tendeloo, Viggo Van; Maurer, Marcus; Campillo-Davò, Diana; Timmermans, Jean‐Pierre; Pintelon, Isabel; Ebo, Didier G.

doi:10.18176/jiaci.0557

Cited by 31 publications

(46 citation statements)

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“…These observations are in line with the results of previous studies with murine cells and human MC lines (11,14,25,26). Our results also confirm that selective MRGPRX2 silencing in primary human MCs via electroporation of DsiRNA against MRGPRX2 enables studying the effects of receptor activation on both calcium mobilization and degranulation (17). As shown previously, this silencing approach has a high transfection efficiency and results in markedly downregulated cell surface levels of MRGPRX2 (17).…”

Section: Discussionsupporting

confidence: 92%

Novel Insights on MRGPRX2-Mediated Hypersensitivity to Neuromuscular Blocking Agents And Fluoroquinolones

et al. 2021

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Neuromuscular blocking agents (NMBAs) like atracurium and rocuronium as well as fluoroquinolones (FQs) cause mast cell-mediated anaphylaxis by activating Mas-related G protein-coupled receptor X2 (MRGPRX2), but many questions remain unanswered. Here, we address three of them, namely whether primary human mast cells show similar activation by these drugs as murine mast cells and mast cell lines, how sugammadex protects from atracurium-induced MRGPRX2-mediated mast cell activation, and why some but not all patients treated with rocuronium develop anaphylaxis. We used peripheral blood-derived cultured mast cells from healthy donors and patients, assessed mast cell activation and degranulation by quantifying intracellular calcium and CD63 expression, respectively, and made use of MRGPRX2-silencing, via electroporation with Dicer-substrate small interfering RNAs, and single cell flow cytometric analyses. Atracurium, ciprofloxacin, and levofloxacin activated and degranulated primary human mast cells, but only MRGPRX2-positive and not MRGPRX2-negative or -silenced mast cells. Sugammadex attenuated the atracurium-induced and MRGPRX2-mediated activation and degranulation of human mast cells by reducing free atracurium levels. The mast cells of patients with IgE-independent anaphylaxis to rocuronium were similar, in their MRGPRX2 expression and function, to those of patients with IgE-mediated anaphylaxis. These findings further improve our understanding of the role and relevance of MRGPRX2-driven mast cell activation in anaphylactic reactions to NMBAs and FQs and may help to improve their prediction, prevention, and treatment.

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Section: Discussionsupporting

confidence: 92%

Novel Insights on MRGPRX2-Mediated Hypersensitivity to Neuromuscular Blocking Agents And Fluoroquinolones

et al. 2021

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“…Many researchers culture human mast cells (HuMC) from CD34 + myeloid progenitor cells derived from buffy coats, cord blood or bone marrow to study human mast cell biology in vitro ( 18 ). These HuMC, which develop from progenitors cells by use of specific culture medium conditions, indeed resemble normal human mast cells closely and have been shown to express MRGPRX2 accordingly ( 19 ). Unfortunately, the method to differentiate them is time consuming and expensive.…”

Section: Introductionmentioning

confidence: 99%

Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2

et al. 2021

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The Mas-related G-protein-coupled receptor X2 (MRGPRX2) is prominently expressed by mast cells and induces degranulation upon binding by different ligands. Its activation has been linked to various mast cell-related diseases, such as chronic spontaneous urticaria, atopic dermatitis and asthma. Therefore, inhibition of MRGPRX2 activity represents a therapeutic target for these conditions. However, the exact pathophysiology of this receptor is still unknown. In vitro research with mast cells is often hampered by the technical limitations of available cell lines. The human mast cell types LAD2 and HuMC (human mast cells cultured from CD34+ progenitor cells) most closely resemble mature human mast cells, yet have a very slow growth rate. A fast proliferating alternative is the human mast cell line HMC1, but they are considered unsuitable for degranulation assays due to their immature phenotype. Moreover, the expression and functionality of MRGPRX2 on HMC1 is controversial. Here, we describe the MRGPRX2 expression and functionality in HMC1 cells, and compare these with LAD2 and HuMC. We also propose a model to render HMC1 suitable for degranulation assays by pre-incubating them with latrunculin-B (Lat-B). Expression of MRGPRX2 by HMC1 was proven by RQ-PCR and flowcytometry, although at lower levels compared with LAD2 and HuMC. Pre-incubation of HMC1 cells with Lat-B significantly increased the overall degranulation capacity, without significantly changing their MRGPRX2 expression, phenotype or morphology. The MRGPRX2 specific compound 48/80 (C48/80) effectively induced degranulation of HMC1 as measured by CD63 membrane expression and β-hexosaminidase release, albeit in lower levels than for LAD2 or HuMC. HMC1, LAD2 and HuMC each had different degranulation kinetics upon stimulation with C48/80. Incubation with the MRGPRX2 specific inhibitor QWF inhibited C48/80-induced degranulation, confirming the functionality of MRGPRX2 on HMC1. In conclusion, HMC1 cells have lower levels of MRGPRX2 expression than LAD2 or HuMC, but are attractive for in vitro research because of their high growth rate and stable phenotype. HMC1 can be used to study MRGPRX2-mediated degranulation after pre-incubation with Lat-B, which provides the opportunity to explore MPRGRX2 biology in mast cells in a feasible way.

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“…2,[5][6][7] None of these studies disclosed MRGPRX2-dependent degranulation, and calcium mobilization was only demonstrated in two of them. 2,7 Actually, the MRGPRX2-dependent MC-activating capacity of rocuronium could well be limited to cutaneous MCs. 4 In the current study, the authors confirm rocuronium to enhance intracellular calcium staining but this effect is not translated in degranulation.…”

Section: O R R E S P O N D E N C E Mrgprx2 Activation In Mast Cells By Neuromuscular Blocking Agents and Other Agonists: Modulation By Sumentioning

confidence: 97%

“…2 Second, at least four studies have explored rocuronium-induced calcium mobilization and degranulation in different cell lines and primary human MCs. 2,[5][6][7] None of these studies disclosed MRGPRX2-dependent degranulation, and calcium mobilization was only demonstrated in two of them. 2,7 Actually, the MRGPRX2-dependent MC-activating capacity of rocuronium could well be limited to cutaneous MCs.…”

Section: O R R E S P O N D E N C E Mrgprx2 Activation In Mast Cells By Neuromuscular Blocking Agents and Other Agonists: Modulation By Sumentioning

confidence: 97%

“…4 In the current study, the authors confirm rocuronium to enhance intracellular calcium staining but this effect is not translated in degranulation. 2,6,7 Hence, our surprise that this rocuronium-sugammadex model was used to further unveil the MRGPRX2-dependent MC degranulating potential of rocuronium. Besides, we have shown that in a majority of our patients, rocuronium hypersensitivity is IgE-mediated.…”

Section: O R R E S P O N D E N C E Mrgprx2 Activation In Mast Cells By Neuromuscular Blocking Agents and Other Agonists: Modulation By Sumentioning

confidence: 99%

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MRGPRX2 activation in mast cells by neuromuscular blocking agents and other agonists: Modulation by Sugammadex by Nithya A. Fernandopulle et al.

Elst

Sabato

Ebo

2021

Clin Experimental Allergy

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MRGPRX2 and Immediate Drug Hypersensitivity: Insights From Cultured Human Mast Cells

Abstract: Peripheral blood cultured mast cells (PBCMCs) are defined as CD117 +ve CD203c +ve cells. PBCMCs harbour two subpopulations: cells with surface expression ofMRGPRX2 (MRGPRX2 +ve) and cells without expression ofMRGPRX2 (MRGPRX2-ve).

Cited by 31 publications

References 0 publications

Novel Insights on MRGPRX2-Mediated Hypersensitivity to Neuromuscular Blocking Agents And Fluoroquinolones

Novel Insights on MRGPRX2-Mediated Hypersensitivity to Neuromuscular Blocking Agents And Fluoroquinolones

Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2

MRGPRX2 activation in mast cells by neuromuscular blocking agents and other agonists: Modulation by Sugammadex by Nithya A. Fernandopulle et al.

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