Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency
Abstract Anti-interferon gamma autoantibodies (anti-IFN-γ autoAbs) neutralize the IFN-γ-mediated functions, contributing to immunodeficiency. A particular autoAb in patient serum had been previously demonstrated to recognize the same determinant on IFN-γ as the neutralizing anti-IFN-γ monoclonal ant...
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-11774-9 |
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| author | Umpa Yasamut Tanchanok Wisitponchai Vannajan Sanghiran Lee Montarop Yamabhai Kuntalee Rangnoi Weeraya Thongkum Koollawat Chupradit Chatchai Tayapiwatana |
| author_facet | Umpa Yasamut Tanchanok Wisitponchai Vannajan Sanghiran Lee Montarop Yamabhai Kuntalee Rangnoi Weeraya Thongkum Koollawat Chupradit Chatchai Tayapiwatana |
| author_sort | Umpa Yasamut |
| collection | DOAJ |
| description | Abstract Anti-interferon gamma autoantibodies (anti-IFN-γ autoAbs) neutralize the IFN-γ-mediated functions, contributing to immunodeficiency. A particular autoAb in patient serum had been previously demonstrated to recognize the same determinant on IFN-γ as the neutralizing anti-IFN-γ monoclonal antibody clone B27 (B27 mAb). This study explored the epitope recognized by B27 mAb. The specific peptide sequence recognized by B27 mAb, TDFLRMMLQEER, was retrieved from a phage display random peptide library. Sequence alignment and homology modeling demonstrated that the queried phage peptide sequence and structure were similar to amino acids at position 27–40 (TLFLGILKNWKEES) of the human IFN-γ. This determinant resides in the contact surface of IFN-γ and interferon gamma receptor 1. To elucidate the crucial amino acids, mutations were introduced by substituting T27 and T27F29L30 with alanine or deleting the amino acid residues T27–L33. The binding of B27 mAb to IFN-γ T27A using western blotting was lesser than that to wild-type. The interaction with triple mutant and T27–L33 deletion mutant using western blotting and sandwich ELISA was abolished. The finding demonstrated that T27, F29, and L30 are critical residues in the B27 antigenic determinant. Identification of the functional domain of IFN-γ decrypted the relevance of neutralizing autoAb in adult-onset immunodeficiency. |
| first_indexed | 2024-12-12T03:18:07Z |
| format | Article |
| id | doaj.art-6cbc1cc7f88144a6b0fbf4b4556b06f3 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-12T03:18:07Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-6cbc1cc7f88144a6b0fbf4b4556b06f32022-12-22T00:40:14ZengNature PortfolioScientific Reports2045-23222022-05-0112111210.1038/s41598-022-11774-9Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiencyUmpa Yasamut0Tanchanok Wisitponchai1Vannajan Sanghiran Lee2Montarop Yamabhai3Kuntalee Rangnoi4Weeraya Thongkum5Koollawat Chupradit6Chatchai Tayapiwatana7Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai UniversityCenter of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai UniversityDepartment of Chemistry, Faculty of Science, University of MalayaMolecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of TechnologyMolecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of TechnologyCenter of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai UniversityCenter of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai UniversityDivision of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai UniversityAbstract Anti-interferon gamma autoantibodies (anti-IFN-γ autoAbs) neutralize the IFN-γ-mediated functions, contributing to immunodeficiency. A particular autoAb in patient serum had been previously demonstrated to recognize the same determinant on IFN-γ as the neutralizing anti-IFN-γ monoclonal antibody clone B27 (B27 mAb). This study explored the epitope recognized by B27 mAb. The specific peptide sequence recognized by B27 mAb, TDFLRMMLQEER, was retrieved from a phage display random peptide library. Sequence alignment and homology modeling demonstrated that the queried phage peptide sequence and structure were similar to amino acids at position 27–40 (TLFLGILKNWKEES) of the human IFN-γ. This determinant resides in the contact surface of IFN-γ and interferon gamma receptor 1. To elucidate the crucial amino acids, mutations were introduced by substituting T27 and T27F29L30 with alanine or deleting the amino acid residues T27–L33. The binding of B27 mAb to IFN-γ T27A using western blotting was lesser than that to wild-type. The interaction with triple mutant and T27–L33 deletion mutant using western blotting and sandwich ELISA was abolished. The finding demonstrated that T27, F29, and L30 are critical residues in the B27 antigenic determinant. Identification of the functional domain of IFN-γ decrypted the relevance of neutralizing autoAb in adult-onset immunodeficiency.https://doi.org/10.1038/s41598-022-11774-9 |
| spellingShingle | Umpa Yasamut Tanchanok Wisitponchai Vannajan Sanghiran Lee Montarop Yamabhai Kuntalee Rangnoi Weeraya Thongkum Koollawat Chupradit Chatchai Tayapiwatana Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency Scientific Reports |
| title | Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency |
| title_full | Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency |
| title_fullStr | Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency |
| title_full_unstemmed | Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency |
| title_short | Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency |
| title_sort | determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency |
| url | https://doi.org/10.1038/s41598-022-11774-9 |
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