Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities
Glycosylation of the immunoglobulin G (IgG)-Fc tail is required for binding to Fc-gamma receptors (FcγRs) and complement-component C1q. A variety of IgG1-glycoforms is detected in human sera. Several groups have found global or antigen-specific skewing of IgG glycosylation, for example in autoimmune...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-08-01
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| Series: | Frontiers in Immunology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fimmu.2017.00877/full |
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| author | Gillian Dekkers Louise Treffers Rosina Plomp Arthur E. H. Bentlage Marcella de Boer Carolien A. M. Koeleman Suzanne N. Lissenberg-Thunnissen Remco Visser Mieke Brouwer Juk Yee Mok Hanke Matlung Timo K. van den Berg Wim J. E. van Esch Taco W. Kuijpers Diana Wouters Theo Rispens Manfred Wuhrer Gestur Vidarsson |
| author_facet | Gillian Dekkers Louise Treffers Rosina Plomp Arthur E. H. Bentlage Marcella de Boer Carolien A. M. Koeleman Suzanne N. Lissenberg-Thunnissen Remco Visser Mieke Brouwer Juk Yee Mok Hanke Matlung Timo K. van den Berg Wim J. E. van Esch Taco W. Kuijpers Diana Wouters Theo Rispens Manfred Wuhrer Gestur Vidarsson |
| author_sort | Gillian Dekkers |
| collection | DOAJ |
| description | Glycosylation of the immunoglobulin G (IgG)-Fc tail is required for binding to Fc-gamma receptors (FcγRs) and complement-component C1q. A variety of IgG1-glycoforms is detected in human sera. Several groups have found global or antigen-specific skewing of IgG glycosylation, for example in autoimmune diseases, viral infections, and alloimmune reactions. The IgG glycoprofiles seem to correlate with disease outcome. Additionally, IgG-glycan composition contributes significantly to Ig-based therapies, as for example IVIg in autoimmune diseases and therapeutic antibodies for cancer treatment. The effect of the different glycan modifications, especially of fucosylation, has been studied before. However, the contribution of the 20 individual IgG glycoforms, in which the combined effect of all 4 modifications, to the IgG function has never been investigated. Here, we combined six glyco-engineering methods to generate all 20 major human IgG1-glycoforms and screened their functional capacity for FcγR and complement activity. Bisection had no effect on FcγR or C1q-binding, and sialylation had no- or little effect on FcγR binding. We confirmed that hypo-fucosylation of IgG1 increased binding to FcγRIIIa and FcγRIIIb by ~17-fold, but in addition we showed that this effect could be further increased to ~40-fold for FcγRIIIa upon simultaneous hypo-fucosylation and hyper-galactosylation, resulting in enhanced NK cell-mediated antibody-dependent cellular cytotoxicity. Moreover, elevated galactosylation and sialylation significantly increased (independent of fucosylation) C1q-binding, downstream complement deposition, and cytotoxicity. In conclusion, fucosylation and galactosylation are primary mediators of functional changes in IgG for FcγR- and complement-mediated effector functions, respectively, with galactose having an auxiliary role for FcγRIII-mediated functions. This knowledge could be used not only for glycan profiling of clinically important (antigen-specific) IgG but also to optimize therapeutic antibody applications. |
| first_indexed | 2024-12-23T13:52:10Z |
| format | Article |
| id | doaj.art-87f60415914a4cbe869393823da2506a |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-23T13:52:10Z |
| publishDate | 2017-08-01 |
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| series | Frontiers in Immunology |
| spelling | doaj.art-87f60415914a4cbe869393823da2506a2022-12-21T17:44:34ZengFrontiers Media S.A.Frontiers in Immunology1664-32242017-08-01810.3389/fimmu.2017.00877279269Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector ActivitiesGillian Dekkers0Louise Treffers1Rosina Plomp2Arthur E. H. Bentlage3Marcella de Boer4Carolien A. M. Koeleman5Suzanne N. Lissenberg-Thunnissen6Remco Visser7Mieke Brouwer8Juk Yee Mok9Hanke Matlung10Timo K. van den Berg11Wim J. E. van Esch12Taco W. Kuijpers13Diana Wouters14Theo Rispens15Manfred Wuhrer16Gestur Vidarsson17Sanquin Research and Landsteiner Laboratory, Department Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Blood Cell Research, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsCenter for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, NetherlandsSanquin Research and Landsteiner Laboratory, Department Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsCenter for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, NetherlandsSanquin Research and Landsteiner Laboratory, Department Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Immunopathology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Reagents, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Blood Cell Research, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Blood Cell Research, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Reagents, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Blood Cell Research, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Immunopathology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsSanquin Research and Landsteiner Laboratory, Department Immunopathology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsCenter for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, NetherlandsSanquin Research and Landsteiner Laboratory, Department Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, NetherlandsGlycosylation of the immunoglobulin G (IgG)-Fc tail is required for binding to Fc-gamma receptors (FcγRs) and complement-component C1q. A variety of IgG1-glycoforms is detected in human sera. Several groups have found global or antigen-specific skewing of IgG glycosylation, for example in autoimmune diseases, viral infections, and alloimmune reactions. The IgG glycoprofiles seem to correlate with disease outcome. Additionally, IgG-glycan composition contributes significantly to Ig-based therapies, as for example IVIg in autoimmune diseases and therapeutic antibodies for cancer treatment. The effect of the different glycan modifications, especially of fucosylation, has been studied before. However, the contribution of the 20 individual IgG glycoforms, in which the combined effect of all 4 modifications, to the IgG function has never been investigated. Here, we combined six glyco-engineering methods to generate all 20 major human IgG1-glycoforms and screened their functional capacity for FcγR and complement activity. Bisection had no effect on FcγR or C1q-binding, and sialylation had no- or little effect on FcγR binding. We confirmed that hypo-fucosylation of IgG1 increased binding to FcγRIIIa and FcγRIIIb by ~17-fold, but in addition we showed that this effect could be further increased to ~40-fold for FcγRIIIa upon simultaneous hypo-fucosylation and hyper-galactosylation, resulting in enhanced NK cell-mediated antibody-dependent cellular cytotoxicity. Moreover, elevated galactosylation and sialylation significantly increased (independent of fucosylation) C1q-binding, downstream complement deposition, and cytotoxicity. In conclusion, fucosylation and galactosylation are primary mediators of functional changes in IgG for FcγR- and complement-mediated effector functions, respectively, with galactose having an auxiliary role for FcγRIII-mediated functions. This knowledge could be used not only for glycan profiling of clinically important (antigen-specific) IgG but also to optimize therapeutic antibody applications.http://journal.frontiersin.org/article/10.3389/fimmu.2017.00877/fullimmunoglobulin G glycosylationFc gamma receptorantibody-dependent cellular cytotoxicitycomplementantibody effector functions |
| spellingShingle | Gillian Dekkers Louise Treffers Rosina Plomp Arthur E. H. Bentlage Marcella de Boer Carolien A. M. Koeleman Suzanne N. Lissenberg-Thunnissen Remco Visser Mieke Brouwer Juk Yee Mok Hanke Matlung Timo K. van den Berg Wim J. E. van Esch Taco W. Kuijpers Diana Wouters Theo Rispens Manfred Wuhrer Gestur Vidarsson Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities Frontiers in Immunology immunoglobulin G glycosylation Fc gamma receptor antibody-dependent cellular cytotoxicity complement antibody effector functions |
| title | Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities |
| title_full | Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities |
| title_fullStr | Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities |
| title_full_unstemmed | Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities |
| title_short | Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities |
| title_sort | decoding the human immunoglobulin g glycan repertoire reveals a spectrum of fc receptor and complement mediated effector activities |
| topic | immunoglobulin G glycosylation Fc gamma receptor antibody-dependent cellular cytotoxicity complement antibody effector functions |
| url | http://journal.frontiersin.org/article/10.3389/fimmu.2017.00877/full |
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