Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency

Yasamut, Umpa and Wisitponchai, Tanchanok and Lee, Vannajan Sanghiran and Yamabhai, Montarop and Rangnoi, Kuntalee and Thongkum, Weeraya and Chupradit, Koollawat and Tayapiwatana, Chatchai (2022) Determination of a distinguished interferon gamma epitope recognized by monoclonal antibody relating to autoantibody associated immunodeficiency. Scientific Reports, 12 (1). ISSN 2045-2322, DOI

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Anti-interferon gamma autoantibodies (anti-IFN-gamma autoAbs) neutralize the IFN-gamma-mediated functions, contributing to immunodeficiency. A particular autoAb in patient serum had been previously demonstrated to recognize the same determinant on IFN-gamma as the neutralizing anti-IFN-gamma 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-gamma. This determinant resides in the contact surface of IFN-gamma 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-gamma 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-gamma decrypted the relevance of neutralizing autoAb in adult-onset immunodeficiency.

Item Type: Article
Funders: Office of the Permanent Secretary, Ministry of Higher Education, Science, Research and Innovation (RGNS 63-067), Distinguished Research Professor Grant (NRCT 808/2563), National Research Council of Thailand (NRCT), Office of National Higher Education Science Research and Innovation Policy Council (NXPO), Thailand, through Program Management Unit for Competitiveness (C10F630145), Thailand Science Research and Innovation (TSRI) (RTA6180012), NRCT (808/2563), SUT Full-time Doctoral Research (61/13/2561)
Uncontrolled Keywords: Distinguished interferon gamma epitope; Monoclonal antibody; Autoantibody associated immunodeficiency
Subjects: Q Science > Q Science (General)
Divisions: Faculty of Science > Department of Chemistry
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 29 Aug 2023 01:47
Last Modified: 29 Aug 2023 01:47

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