論文 - 詳細
| RRC ID | 50081 |
|---|---|
| 著者 | Watanabe N, Suzuki Y, Yonezu T, Nakagawa Y, Shiina T, Hirayama N, Inokuchi S, Inoue S. |
| タイトル | A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule. |
| ジャーナル | Sci Rep |
| 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 DRA*01:01 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 (DRB1*01:01) 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. |
| 巻・号 | 7(1) |
| ページ | 6798 |
| 公開日 | 2017-7-28 |
| DOI | 10.1038/s41598-017-07080-4 |
| PII | 10.1038/s41598-017-07080-4 |
| PMID | 28754892 |
| PMC | PMC5533769 |
| MeSH | 3T3 Cells Animals Antigen Presentation Autoantigens / immunology* HEK293 Cells HLA-DRB1 Chains / immunology* High-Throughput Screening Assays / methods* Humans Mice Myelin Basic Protein / immunology Peptides / immunology |
| IF | 3.998 |
| 引用数 | 2 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 3 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 遺伝子材料 | pCAG-HIVgp (RDB04394) pCMV-VSV-G-RSV-Rev (RDB04393). |