Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panel
Abstract Fc‐gamma receptor (FcγR) activation by soluble IgG immune complexes (sICs) represents a major mechanism of inflammation in certain autoimmune diseases such as systemic lupus erythematosus (SLE). A robust and scalable test system allowing for the detection and quantification of sIC bioactivi...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Springer Nature
2021-11-01
|
| Series: | EMBO Molecular Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.15252/emmm.202114182 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849761565577838592 |
|---|---|
| author | Haizhang Chen Andrea Maul‐Pavicic Martin Holzer Magdalena Huber Ulrich Salzer Nina Chevalier Reinhard E Voll Hartmut Hengel Philipp Kolb |
| author_facet | Haizhang Chen Andrea Maul‐Pavicic Martin Holzer Magdalena Huber Ulrich Salzer Nina Chevalier Reinhard E Voll Hartmut Hengel Philipp Kolb |
| author_sort | Haizhang Chen |
| collection | DOAJ |
| description | Abstract Fc‐gamma receptor (FcγR) activation by soluble IgG immune complexes (sICs) represents a major mechanism of inflammation in certain autoimmune diseases such as systemic lupus erythematosus (SLE). A robust and scalable test system allowing for the detection and quantification of sIC bioactivity is missing. We developed a comprehensive reporter cell panel detecting activation of FcγRs. The reporter cell lines were integrated into an assay that enables the quantification of sIC reactivity via ELISA or a faster detection using flow cytometry. This identified FcγRIIA(H) and FcγRIIIA as the most sIC‐sensitive FcγRs in our test system. Reaching a detection limit in the very low nanomolar range, the assay proved also to be sensitive to sIC stoichiometry and size reproducing for the first time a complete Heidelberger‐Kendall curve in terms of immune receptor activation. Analyzing sera from SLE patients and mouse models of lupus and arthritis proved that sIC‐dependent FcγR activation has predictive capabilities regarding severity of SLE disease. The assay provides a sensitive and scalable tool to evaluate the size, amount, and bioactivity of sICs in all settings. |
| format | Article |
| id | doaj-art-d3c81898e97a42de975a99761a93314a |
| institution | DOAJ |
| issn | 1757-4676 1757-4684 |
| language | English |
| publishDate | 2021-11-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | EMBO Molecular Medicine |
| spelling | doaj-art-d3c81898e97a42de975a99761a93314a2025-08-20T03:06:00ZengSpringer NatureEMBO Molecular Medicine1757-46761757-46842021-11-0114111710.15252/emmm.202114182Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panelHaizhang Chen0Andrea Maul‐Pavicic1Martin Holzer2Magdalena Huber3Ulrich Salzer4Nina Chevalier5Reinhard E Voll6Hartmut Hengel7Philipp Kolb8Institute of Virology, University Medical Center, Albert‐Ludwigs‐University FreiburgDepartment of Rheumatology and Clinical Immunology, Medical Center – University of Freiburg, Faculty of Medicine, University of FreiburgInstitute for Pharmaceutical Sciences, Albert‐Ludwigs‐University FreiburgInstitute of Virology, University Medical Center, Albert‐Ludwigs‐University FreiburgDepartment of Rheumatology and Clinical Immunology, Medical Center – University of Freiburg, Faculty of Medicine, University of FreiburgDepartment of Rheumatology and Clinical Immunology, Medical Center – University of Freiburg, Faculty of Medicine, University of FreiburgDepartment of Rheumatology and Clinical Immunology, Medical Center – University of Freiburg, Faculty of Medicine, University of FreiburgInstitute of Virology, University Medical Center, Albert‐Ludwigs‐University FreiburgInstitute of Virology, University Medical Center, Albert‐Ludwigs‐University FreiburgAbstract Fc‐gamma receptor (FcγR) activation by soluble IgG immune complexes (sICs) represents a major mechanism of inflammation in certain autoimmune diseases such as systemic lupus erythematosus (SLE). A robust and scalable test system allowing for the detection and quantification of sIC bioactivity is missing. We developed a comprehensive reporter cell panel detecting activation of FcγRs. The reporter cell lines were integrated into an assay that enables the quantification of sIC reactivity via ELISA or a faster detection using flow cytometry. This identified FcγRIIA(H) and FcγRIIIA as the most sIC‐sensitive FcγRs in our test system. Reaching a detection limit in the very low nanomolar range, the assay proved also to be sensitive to sIC stoichiometry and size reproducing for the first time a complete Heidelberger‐Kendall curve in terms of immune receptor activation. Analyzing sera from SLE patients and mouse models of lupus and arthritis proved that sIC‐dependent FcγR activation has predictive capabilities regarding severity of SLE disease. The assay provides a sensitive and scalable tool to evaluate the size, amount, and bioactivity of sICs in all settings.https://doi.org/10.15252/emmm.202114182immune complexesFc‐gamma receptorsFcγR activationSLE |
| spellingShingle | Haizhang Chen Andrea Maul‐Pavicic Martin Holzer Magdalena Huber Ulrich Salzer Nina Chevalier Reinhard E Voll Hartmut Hengel Philipp Kolb Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panel EMBO Molecular Medicine immune complexes Fc‐gamma receptors FcγR activation SLE |
| title | Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panel |
| title_full | Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panel |
| title_fullStr | Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panel |
| title_full_unstemmed | Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panel |
| title_short | Detection and functional resolution of soluble immune complexes by an FcγR reporter cell panel |
| title_sort | detection and functional resolution of soluble immune complexes by an fcγr reporter cell panel |
| topic | immune complexes Fc‐gamma receptors FcγR activation SLE |
| url | https://doi.org/10.15252/emmm.202114182 |
| work_keys_str_mv | AT haizhangchen detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT andreamaulpavicic detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT martinholzer detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT magdalenahuber detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT ulrichsalzer detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT ninachevalier detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT reinhardevoll detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT hartmuthengel detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel AT philippkolb detectionandfunctionalresolutionofsolubleimmunecomplexesbyanfcgrreportercellpanel |