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BRIEF RESEARCH REPORT article

Front. Immunol.
Sec. Molecular Innate Immunity
Volume 15 - 2024 | doi: 10.3389/fimmu.2024.1506034

Functional MRGPRX2 expression on peripheral blood-derived human mast cells increases at low seeding density and is suppressed by interleukin-9 and fetal bovine serum Authors

Provisionally accepted
Toon Ieven Toon Ieven 1,2*Janne Goossens Janne Goossens 1Willem Roosens Willem Roosens 1,2Anne-Charlotte Jonckheere Anne-Charlotte Jonckheere 1Jonathan Cremer Jonathan Cremer 1Ellen Dilissen Ellen Dilissen 1Rune Persoons Rune Persoons 1Lieven Dupont Lieven Dupont 3,4Rik Schrijvers Rik Schrijvers 1,2Peter Vandenberghe Peter Vandenberghe 5,6Christine Breynaert Christine Breynaert 1,2Dominique MA Bullens Dominique MA Bullens 1,7*
  • 1 KU Leuven Department of Microbiology, Immunology and Transplantation, Allergy and Clinical Immunology Research Group, Leuven, Belgium
  • 2 Division of General Internal Medicine, Allergy and Clinical Immunology, UZ Leuven, Leuven, Belgium
  • 3 KU Leuven Department of Chronic Diseases and Metabolism, Laboratory of Respiratory Diseases and Thoracic Surgery (BREATHE), Leuven, Belgium
  • 4 Division of Respiratory Diseases, UZ Leuven, Leuven, Belgium
  • 5 KU Leuven Department of Human Genetics, Laboratory for Genetics of Malignant Disorders, Leuven, Belgium
  • 6 Division of Hematology, UZ Leuven, Leuven, Belgium
  • 7 Division of Pediatrics, UZ Leuven, Leuven, Belgium

The final, formatted version of the article will be published soon.

    Primary human mast cells (MC) obtained through culturing of blood-derived MC progenitors are the preferred model for the ex vivo study of MRGPRX2-vs. IgE-mediated MC activation. In order to assess the impact of culture conditions on functional MRGPRX2 expression, we cultured CD34+ -enriched PBMC from peripheral whole blood (PB) and buffy coat (BC) samples in MethoCult medium containing stem cell factor (SCF) and interleukin (IL)-3, modified through variations in seeding density and adding or withholding IL-6, IL-9 and fetal bovine serum (FBS). Functional expression of MRGPRX2 was assessed after 4 weeks via flow cytometry. We found similar proportions of CD34 + MC-committed progenitors in BC and PB. Higher seeding densities (≥ 1x10 5 cells/mL) and exposure to IL-9 and FBS suppressed functional MRGPRX2 expression at 4 weeks, while leaving MC yield largely unaffected. IL-6 had no impact on MRGPRX2 expression. MRGPRX2-expressing MC upregulated CD63 upon stimulation with polyclonal anti-IgE, substance P and compound 48/80 at 4 weeks. Ketotifen and dasatinib but not cromolyn sodium inhibited both IgE-and MRGPRX2-dependent pathways. Our results confirm the feasibility of functional MC activation studies on PB-derived MC after a short 4-week culture and highlight the impact of culture conditions on functional MRGPRX2 expression.

    Keywords: (8/8) human, Blood, mast cell, culture, MRGPRX2, Interleukin-9, Serum, Ketotifen

    Received: 04 Oct 2024; Accepted: 02 Dec 2024.

    Copyright: © 2024 Ieven, Goossens, Roosens, Jonckheere, Cremer, Dilissen, Persoons, Dupont, Schrijvers, Vandenberghe, Breynaert and Bullens. 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) or licensor 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.

    * Correspondence:
    Toon Ieven, KU Leuven Department of Microbiology, Immunology and Transplantation, Allergy and Clinical Immunology Research Group, Leuven, Belgium
    Dominique MA Bullens, KU Leuven Department of Microbiology, Immunology and Transplantation, Allergy and Clinical Immunology Research Group, Leuven, Belgium

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.