Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodies
IntroductionAlthough Interleukin (IL)-6-type cytokine signaling is critical for maintaining the body's homeostasis, aberrant signaling has been observed in numerous diseases including autoimmunity and cancer. Currently, all approved biologics that inhibit IL-6-type cytokines specifically target...
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Frontiers Media S.A.
2025-08-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1613004/full |
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| author | Silke Pudewell Julia Heuser Inna Dorogobed Britta Lipinski Thi Hong Hue Tran Pia Metzenmacher Richard Kunze Felix Geyer Stefan Zielonka Doreen M. Floss Harald Kolmar Jens M. Moll Jürgen Scheller |
| author_facet | Silke Pudewell Julia Heuser Inna Dorogobed Britta Lipinski Thi Hong Hue Tran Pia Metzenmacher Richard Kunze Felix Geyer Stefan Zielonka Doreen M. Floss Harald Kolmar Jens M. Moll Jürgen Scheller |
| author_sort | Silke Pudewell |
| collection | DOAJ |
| description | IntroductionAlthough Interleukin (IL)-6-type cytokine signaling is critical for maintaining the body's homeostasis, aberrant signaling has been observed in numerous diseases including autoimmunity and cancer. Currently, all approved biologics that inhibit IL-6-type cytokines specifically target the key pro-inflammatory mediator IL-6 or its receptor (IL6R). Historically, direct inhibition of glycoprotein 130 (gp130)—the shared transmembrane receptor for IL-6-type cytokines—was avoided due to concerns that broad suppression might cause more harm than benefit. However, this view is being reconsidered in light of the clinical success of Janus kinase (JAK) inhibitors, which broadly disrupt cytokine signaling, including pathways mediated by gp130.MethodsHere we developed four single domain antibodies (sdAb), consisting out of a camelid-derived nanobody and a human Fc-fragment, and characterized them by direct protein interaction analysis, epitope binding, epitope binning, as well as inhibition of cytokine-induced stimulation and proliferation of appropriate Ba/F3 cell lines and trans-migration in HT-29 cells.ResultsThe four sdAb-Fc constructs GP01-, GP11- GP13- and GP20-Fc bind directly to gp130 in the cytokine binding module (CBM) and largely inhibit IL-6-type cytokine signaling by interfering with the high-affinity binding site of IL-6, IL-11, CLCF1, CT1, CNTF, OSM and LIF. Furthermore, we functionally demonstrate the inhibitory effect of the selected nanobodies in cell-based transmigration assays of the human colorectal cancer cell line HT-29.DiscussionIn summary, our study has identified and characterized four novel inhibitory high-affinity gp130 nanobodies with potential for use in cytokine-dependent autoimmunity or cancer therapy. |
| format | Article |
| id | doaj-art-59b3eee090e8459db916258a3399a259 |
| institution | Kabale University |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj-art-59b3eee090e8459db916258a3399a2592025-08-20T04:01:00ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-08-011610.3389/fimmu.2025.16130041613004Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodiesSilke Pudewell0Julia Heuser1Inna Dorogobed2Britta Lipinski3Thi Hong Hue Tran4Pia Metzenmacher5Richard Kunze6Felix Geyer7Stefan Zielonka8Doreen M. Floss9Harald Kolmar10Jens M. Moll11Jürgen Scheller12Institute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, GermanyApplied Biochemistry, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, GermanyBiomolecular Immunotherapy, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, GermanyBiomolecular Immunotherapy, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, GermanyApplied Biochemistry, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, GermanyBiomolecular Immunotherapy, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, GermanyApplied Biochemistry, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, GermanyIntroductionAlthough Interleukin (IL)-6-type cytokine signaling is critical for maintaining the body's homeostasis, aberrant signaling has been observed in numerous diseases including autoimmunity and cancer. Currently, all approved biologics that inhibit IL-6-type cytokines specifically target the key pro-inflammatory mediator IL-6 or its receptor (IL6R). Historically, direct inhibition of glycoprotein 130 (gp130)—the shared transmembrane receptor for IL-6-type cytokines—was avoided due to concerns that broad suppression might cause more harm than benefit. However, this view is being reconsidered in light of the clinical success of Janus kinase (JAK) inhibitors, which broadly disrupt cytokine signaling, including pathways mediated by gp130.MethodsHere we developed four single domain antibodies (sdAb), consisting out of a camelid-derived nanobody and a human Fc-fragment, and characterized them by direct protein interaction analysis, epitope binding, epitope binning, as well as inhibition of cytokine-induced stimulation and proliferation of appropriate Ba/F3 cell lines and trans-migration in HT-29 cells.ResultsThe four sdAb-Fc constructs GP01-, GP11- GP13- and GP20-Fc bind directly to gp130 in the cytokine binding module (CBM) and largely inhibit IL-6-type cytokine signaling by interfering with the high-affinity binding site of IL-6, IL-11, CLCF1, CT1, CNTF, OSM and LIF. Furthermore, we functionally demonstrate the inhibitory effect of the selected nanobodies in cell-based transmigration assays of the human colorectal cancer cell line HT-29.DiscussionIn summary, our study has identified and characterized four novel inhibitory high-affinity gp130 nanobodies with potential for use in cytokine-dependent autoimmunity or cancer therapy.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1613004/fullIL-6-type cytokinesglycoprotein 130single domain antibodyinhibitorinflammation |
| spellingShingle | Silke Pudewell Julia Heuser Inna Dorogobed Britta Lipinski Thi Hong Hue Tran Pia Metzenmacher Richard Kunze Felix Geyer Stefan Zielonka Doreen M. Floss Harald Kolmar Jens M. Moll Jürgen Scheller Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodies Frontiers in Immunology IL-6-type cytokines glycoprotein 130 single domain antibody inhibitor inflammation |
| title | Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodies |
| title_full | Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodies |
| title_fullStr | Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodies |
| title_full_unstemmed | Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodies |
| title_short | Targeting the major pro-inflammatory interleukin-6-type cytokine receptor gp130 by antagonistic single domain antibodies |
| title_sort | targeting the major pro inflammatory interleukin 6 type cytokine receptor gp130 by antagonistic single domain antibodies |
| topic | IL-6-type cytokines glycoprotein 130 single domain antibody inhibitor inflammation |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1613004/full |
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