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

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. There...

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Autores principales: Hermans, Maud A. W., van Stigt, Astrid C., van de Meerendonk, Sanne, Schrijver, Benjamin, van Daele, Paul L. A., van Hagen, Petrus M., van Splunter, Marloes, Dik, Willem A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8006456/
https://www.ncbi.nlm.nih.gov/pubmed/33790895
http://dx.doi.org/10.3389/fimmu.2021.625284
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author Hermans, Maud A. W.
van Stigt, Astrid C.
van de Meerendonk, Sanne
Schrijver, Benjamin
van Daele, Paul L. A.
van Hagen, Petrus M.
van Splunter, Marloes
Dik, Willem A.
author_facet Hermans, Maud A. W.
van Stigt, Astrid C.
van de Meerendonk, Sanne
Schrijver, Benjamin
van Daele, Paul L. A.
van Hagen, Petrus M.
van Splunter, Marloes
Dik, Willem A.
author_sort Hermans, Maud A. W.
collection PubMed
description 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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spelling pubmed-80064562021-03-30 Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2 Hermans, Maud A. W. van Stigt, Astrid C. van de Meerendonk, Sanne Schrijver, Benjamin van Daele, Paul L. A. van Hagen, Petrus M. van Splunter, Marloes Dik, Willem A. Front Immunol Immunology 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. Frontiers Media S.A. 2021-03-15 /pmc/articles/PMC8006456/ /pubmed/33790895 http://dx.doi.org/10.3389/fimmu.2021.625284 Text en Copyright © 2021 Hermans, van Stigt, van de Meerendonk, Schrijver, van Daele, van Hagen, van Splunter and Dik http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Hermans, Maud A. W.
van Stigt, Astrid C.
van de Meerendonk, Sanne
Schrijver, Benjamin
van Daele, Paul L. A.
van Hagen, Petrus M.
van Splunter, Marloes
Dik, Willem A.
Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2
title Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2
title_full Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2
title_fullStr Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2
title_full_unstemmed Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2
title_short Human Mast Cell Line HMC1 Expresses Functional Mas-Related G-Protein Coupled Receptor 2
title_sort human mast cell line hmc1 expresses functional mas-related g-protein coupled receptor 2
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8006456/
https://www.ncbi.nlm.nih.gov/pubmed/33790895
http://dx.doi.org/10.3389/fimmu.2021.625284
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