A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule

Abstract A number of autoimmune diseases are associated with the genotypes of human leukocyte antigen class II (HLA), some of which present peptides derived from self-proteins, resulting in clonal expansion of self-reactive T cells. Therefore, selective inhibition of self-peptide loading onto such d...

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Autores principales:	Nobuo Watanabe, Yusuke Suzuki, Takahisa Yonezu, Yuki Nakagawa, Takashi Shiina, Noriaki Hirayama, Sadaki Inokuchi, Shigeaki Inoue
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a45b10110a54476c84daacb6b46f2609
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spelling	oai:doaj.org-article:a45b10110a54476c84daacb6b46f26092021-12-02T11:40:13ZA cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule10.1038/s41598-017-07080-42045-2322https://doaj.org/article/a45b10110a54476c84daacb6b46f26092017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-07080-4https://doaj.org/toc/2045-2322Abstract A number of autoimmune diseases are associated with the genotypes of human leukocyte antigen class II (HLA), some of which present peptides derived from self-proteins, resulting in clonal expansion of self-reactive T cells. Therefore, selective inhibition of self-peptide loading onto such disease-associated HLA could ameliorate the diseases. To effectively identify such compounds, in this study, we established, for the first time, a cell- and 96-well microplate-based high-throughput screening system for inhibitors of antigen presentation. A panel of DRB1 genes plus DRA01:01 gene were expressed in HEK293T cells and in 3T3 cells, and their binding with biotinylated known self-antigen peptides was measured by flow cytometry. HLA-DR1 (DRB101:01) and DR15 (DRB1*15:01) showed a high affinity with myelin basic protein peptide (MBP83-98). Therefore, in 96-well plate wells, MBP83-99 was allowed to bind to DR1 or DR15 on 3T3 cells in competition with a test compound, and the HLA-bound peptide was detected by streptavidin-conjugated β-galactosidase, thereby identifying inhibitor compounds for rheumatoid arthritis or multiple sclerosis. Our assay system has a potential for broad applications, including designing peptide vaccines.Nobuo WatanabeYusuke SuzukiTakahisa YonezuYuki NakagawaTakashi ShiinaNoriaki HirayamaSadaki InokuchiShigeaki InoueNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nobuo Watanabe Yusuke Suzuki Takahisa Yonezu Yuki Nakagawa Takashi Shiina Noriaki Hirayama Sadaki Inokuchi Shigeaki Inoue A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule
description	Abstract A number of autoimmune diseases are associated with the genotypes of human leukocyte antigen class II (HLA), some of which present peptides derived from self-proteins, resulting in clonal expansion of self-reactive T cells. Therefore, selective inhibition of self-peptide loading onto such disease-associated HLA could ameliorate the diseases. To effectively identify such compounds, in this study, we established, for the first time, a cell- and 96-well microplate-based high-throughput screening system for inhibitors of antigen presentation. A panel of DRB1 genes plus DRA01:01 gene were expressed in HEK293T cells and in 3T3 cells, and their binding with biotinylated known self-antigen peptides was measured by flow cytometry. HLA-DR1 (DRB101:01) and DR15 (DRB1*15:01) showed a high affinity with myelin basic protein peptide (MBP83-98). Therefore, in 96-well plate wells, MBP83-99 was allowed to bind to DR1 or DR15 on 3T3 cells in competition with a test compound, and the HLA-bound peptide was detected by streptavidin-conjugated β-galactosidase, thereby identifying inhibitor compounds for rheumatoid arthritis or multiple sclerosis. Our assay system has a potential for broad applications, including designing peptide vaccines.
format	article
author	Nobuo Watanabe Yusuke Suzuki Takahisa Yonezu Yuki Nakagawa Takashi Shiina Noriaki Hirayama Sadaki Inokuchi Shigeaki Inoue
author_facet	Nobuo Watanabe Yusuke Suzuki Takahisa Yonezu Yuki Nakagawa Takashi Shiina Noriaki Hirayama Sadaki Inokuchi Shigeaki Inoue
author_sort	Nobuo Watanabe
title	A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule
title_short	A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule
title_full	A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule
title_fullStr	A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule
title_full_unstemmed	A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule
title_sort	cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by hla class ii molecule
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/a45b10110a54476c84daacb6b46f2609
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A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule

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