Human Fcγ-receptors selectively respond to C-reactive protein isoforms

IntroductionThe pentameric C-reactive protein (pCRP), an acute-phase protein, binds to lysophosphatidylcholine (LPC) displayed on the surface of dying cells and microorganisms to activate the complement system and to opsonize immune cells via Fcγ-receptors (FcγRs). Members of the FcγR family are cha...

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Main Authors: Anna Henning, Johanna Seer, Johannes Zeller, Karlheinz Peter, Haizhang Chen, Julia Thomé, Philipp Kolb, Steffen U. Eisenhardt, Katja Hoffmann, Hartmut Hengel
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1598605/full
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author Anna Henning
Johanna Seer
Johannes Zeller
Karlheinz Peter
Karlheinz Peter
Haizhang Chen
Julia Thomé
Philipp Kolb
Steffen U. Eisenhardt
Katja Hoffmann
Hartmut Hengel
author_facet Anna Henning
Johanna Seer
Johannes Zeller
Karlheinz Peter
Karlheinz Peter
Haizhang Chen
Julia Thomé
Philipp Kolb
Steffen U. Eisenhardt
Katja Hoffmann
Hartmut Hengel
author_sort Anna Henning
collection DOAJ
description IntroductionThe pentameric C-reactive protein (pCRP), an acute-phase protein, binds to lysophosphatidylcholine (LPC) displayed on the surface of dying cells and microorganisms to activate the complement system and to opsonize immune cells via Fcγ-receptors (FcγRs). Members of the FcγR family are characterized by the recognition of the Fc part of IgG antibodies.MethodsWe utilized a mouse thymoma BW5147 reporter cell panel stably expressing chimeric human FcγR-CD3ζ-chain receptors to define the molecular requirements for FcγR crosslinking by C-reactive protein (CRP).ResultsApplying this approach, we show a robust activation of CD64/FcγRI and CD32a/FcγRIIa by immobilized CRP isoforms as well as triggering of inhibitory CD32b/FcγRIIb. Of note, activation of FcγRIIa was restricted to the 131R allelic variant but not observed with 131H. In contrast, FcγRIII isoforms CD16aF, CD16aV and CD16b were not activated by pCRP, although binding of CRP isoforms to FcγRIII was detectable. Activation of FcγRs by free pCRP in solution phase was considerably lower than with immobilized pCRP on hydrophilic plastic surfaces and readily abolished by IgG at serum level concentrations, whereas it was enhanced by the addition of streptococci. The types of FcγRs mainly responding to pCRP in solution phase (CD64/FcγRI and CD32aR/FcγRIIaR) clearly differed from FcγRs responding to soluble multimeric IgG complexes (i.e., CD16aV/FcγRIIIaV and CD32aH/FcγRIIaH). Compared to pCRP, monomeric CRP (mCRP) showed lower levels of activation in those selective FcγRs. FcγR activation was linked to recognition by conformation-dependent CRP antibodies. Unmasking of the mAb 9C9-defined neoepitope in pCRP* correlated with the triggering of FcγRs, indicating that pCRP* is the major FcγR-activating CRP conformation.DiscussionThe assay provides a novel, scalable approach to determine the molecular properties of CRP as a physiological ligand of FcγR-mediated bioactivities.
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spelling doaj-art-62f5c4babafa44cf8c3413e8ddf5baa72025-08-20T03:53:46ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-05-011610.3389/fimmu.2025.15986051598605Human Fcγ-receptors selectively respond to C-reactive protein isoformsAnna Henning0Johanna Seer1Johannes Zeller2Karlheinz Peter3Karlheinz Peter4Haizhang Chen5Julia Thomé6Philipp Kolb7Steffen U. Eisenhardt8Katja Hoffmann9Hartmut Hengel10Institute of Virology, University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, GermanyInstitute of Virology, University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, GermanyDepartment of Plastic and Hand Surgery, University of Freiburg Medical Centre, Medical Faculty of the University of Freiburg, Freiburg, GermanyBaker Department of Cardiometabolic Health, University of Melbourne, Parkville, VIC, AustraliaAtherothrombosis and Vascular Biology Laboratory, Baker Heart and Diabetes Institute, Melbourne, VIC, AustraliaInstitute of Virology, University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, GermanyDepartment of Plastic and Hand Surgery, University of Freiburg Medical Centre, Medical Faculty of the University of Freiburg, Freiburg, GermanyInstitute of Virology, University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, GermanyDepartment of Plastic and Hand Surgery, University of Freiburg Medical Centre, Medical Faculty of the University of Freiburg, Freiburg, GermanyInstitute of Virology, University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, GermanyInstitute of Virology, University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, GermanyIntroductionThe pentameric C-reactive protein (pCRP), an acute-phase protein, binds to lysophosphatidylcholine (LPC) displayed on the surface of dying cells and microorganisms to activate the complement system and to opsonize immune cells via Fcγ-receptors (FcγRs). Members of the FcγR family are characterized by the recognition of the Fc part of IgG antibodies.MethodsWe utilized a mouse thymoma BW5147 reporter cell panel stably expressing chimeric human FcγR-CD3ζ-chain receptors to define the molecular requirements for FcγR crosslinking by C-reactive protein (CRP).ResultsApplying this approach, we show a robust activation of CD64/FcγRI and CD32a/FcγRIIa by immobilized CRP isoforms as well as triggering of inhibitory CD32b/FcγRIIb. Of note, activation of FcγRIIa was restricted to the 131R allelic variant but not observed with 131H. In contrast, FcγRIII isoforms CD16aF, CD16aV and CD16b were not activated by pCRP, although binding of CRP isoforms to FcγRIII was detectable. Activation of FcγRs by free pCRP in solution phase was considerably lower than with immobilized pCRP on hydrophilic plastic surfaces and readily abolished by IgG at serum level concentrations, whereas it was enhanced by the addition of streptococci. The types of FcγRs mainly responding to pCRP in solution phase (CD64/FcγRI and CD32aR/FcγRIIaR) clearly differed from FcγRs responding to soluble multimeric IgG complexes (i.e., CD16aV/FcγRIIIaV and CD32aH/FcγRIIaH). Compared to pCRP, monomeric CRP (mCRP) showed lower levels of activation in those selective FcγRs. FcγR activation was linked to recognition by conformation-dependent CRP antibodies. Unmasking of the mAb 9C9-defined neoepitope in pCRP* correlated with the triggering of FcγRs, indicating that pCRP* is the major FcγR-activating CRP conformation.DiscussionThe assay provides a novel, scalable approach to determine the molecular properties of CRP as a physiological ligand of FcγR-mediated bioactivities.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1598605/fullC-reactive proteinCRP isoformsFcγ receptorimmunoglobulinimmune complex
spellingShingle Anna Henning
Johanna Seer
Johannes Zeller
Karlheinz Peter
Karlheinz Peter
Haizhang Chen
Julia Thomé
Philipp Kolb
Steffen U. Eisenhardt
Katja Hoffmann
Hartmut Hengel
Human Fcγ-receptors selectively respond to C-reactive protein isoforms
Frontiers in Immunology
C-reactive protein
CRP isoforms
Fcγ receptor
immunoglobulin
immune complex
title Human Fcγ-receptors selectively respond to C-reactive protein isoforms
title_full Human Fcγ-receptors selectively respond to C-reactive protein isoforms
title_fullStr Human Fcγ-receptors selectively respond to C-reactive protein isoforms
title_full_unstemmed Human Fcγ-receptors selectively respond to C-reactive protein isoforms
title_short Human Fcγ-receptors selectively respond to C-reactive protein isoforms
title_sort human fcγ receptors selectively respond to c reactive protein isoforms
topic C-reactive protein
CRP isoforms
Fcγ receptor
immunoglobulin
immune complex
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1598605/full
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