Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B Cells
This study reports the use of single-cell RNA sequencing to evaluate B cells in the peripheral blood mononuclear cells (PBMCs) and intrathyroidal blood mononuclear cells of patients with Graves’ disease (GD) undergoing thyroidectomy. These cells were stimulated with overlapping peptides of thyroid a...
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2025-07-01
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| author | Genki Kobayashi Takuro Okamura Yoshitaka Hashimoto Kimiko Sakai Madoka Sumi Dan Imai Nobuko Kitagawa Masahide Hamaguchi Michiaki Fukui |
| author_facet | Genki Kobayashi Takuro Okamura Yoshitaka Hashimoto Kimiko Sakai Madoka Sumi Dan Imai Nobuko Kitagawa Masahide Hamaguchi Michiaki Fukui |
| author_sort | Genki Kobayashi |
| collection | DOAJ |
| description | This study reports the use of single-cell RNA sequencing to evaluate B cells in the peripheral blood mononuclear cells (PBMCs) and intrathyroidal blood mononuclear cells of patients with Graves’ disease (GD) undergoing thyroidectomy. These cells were stimulated with overlapping peptides of thyroid autoantigens, including thyroid-stimulating hormone receptor (TSHR), thyroglobulin (Tg), and thyroid peroxidase (TPO). In PBMCs, naive B cells are characterized by <i>IL6</i> and <i>CXCR5</i>, whereas memory B cells express <i>IGHG1</i>, <i>IGHG2</i>, and <i>CD74</i>. <i>HLA-DMA</i>, <i>HLA-DRB1</i>, <i>IGHG</i>, <i>IGHM</i>, <i>CD74</i>, <i>CD79A</i>, and <i>MS4A1</i> expression increased in peptide-stimulated naive and memory B cells compared to those in the controls. Thyroid naive B cells are characterized by <i>CD40</i> and <i>TNFRSF13C</i>, whereas memory B cells express <i>IGHM</i>, <i>CD79A</i>, and <i>MS4A1.</i> Thyroid B cells showed higher <i>DUSP1</i>, <i>DUSP2</i>, <i>CD69</i>, <i>FOSB</i>, <i>RGS1</i>, and immunoglobulin gene expression than control PBMCs and thyroid cells. B-cell receptor analysis revealed frequent <i>IGHV3-23</i> and <i>IGHV4-34</i> usage in controls, whereas <i>IGHV4-34/IGHJ4</i> expression was increased in TSHR-stimulated groups. We concluded that B-cell responses to TSHR, Tg, and TPO differed and that changes in B-cell reactivity also occurred in PBMCs and the thyroid. Additionally, <i>IGHV3-23</i> and <i>IGHV4-34</i> may be associated with autoantibody production in GD. |
| format | Article |
| id | doaj-art-b84d5ad3ddbd4de68d79fbbe838f88c5 |
| institution | Kabale University |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-07-01 |
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| spelling | doaj-art-b84d5ad3ddbd4de68d79fbbe838f88c52025-08-20T03:36:15ZengMDPI AGCells2073-44092025-07-011414110210.3390/cells14141102Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B CellsGenki Kobayashi0Takuro Okamura1Yoshitaka Hashimoto2Kimiko Sakai3Madoka Sumi4Dan Imai5Nobuko Kitagawa6Masahide Hamaguchi7Michiaki Fukui8Department of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanDepartment of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanDepartment of Diabetes and Endocrinology, Matsushita Memorial Hospital, Moriguchi 570-8540, JapanDepartment of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanDepartment of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanDepartment of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanDepartment of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanDepartment of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanDepartment of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, JapanThis study reports the use of single-cell RNA sequencing to evaluate B cells in the peripheral blood mononuclear cells (PBMCs) and intrathyroidal blood mononuclear cells of patients with Graves’ disease (GD) undergoing thyroidectomy. These cells were stimulated with overlapping peptides of thyroid autoantigens, including thyroid-stimulating hormone receptor (TSHR), thyroglobulin (Tg), and thyroid peroxidase (TPO). In PBMCs, naive B cells are characterized by <i>IL6</i> and <i>CXCR5</i>, whereas memory B cells express <i>IGHG1</i>, <i>IGHG2</i>, and <i>CD74</i>. <i>HLA-DMA</i>, <i>HLA-DRB1</i>, <i>IGHG</i>, <i>IGHM</i>, <i>CD74</i>, <i>CD79A</i>, and <i>MS4A1</i> expression increased in peptide-stimulated naive and memory B cells compared to those in the controls. Thyroid naive B cells are characterized by <i>CD40</i> and <i>TNFRSF13C</i>, whereas memory B cells express <i>IGHM</i>, <i>CD79A</i>, and <i>MS4A1.</i> Thyroid B cells showed higher <i>DUSP1</i>, <i>DUSP2</i>, <i>CD69</i>, <i>FOSB</i>, <i>RGS1</i>, and immunoglobulin gene expression than control PBMCs and thyroid cells. B-cell receptor analysis revealed frequent <i>IGHV3-23</i> and <i>IGHV4-34</i> usage in controls, whereas <i>IGHV4-34/IGHJ4</i> expression was increased in TSHR-stimulated groups. We concluded that B-cell responses to TSHR, Tg, and TPO differed and that changes in B-cell reactivity also occurred in PBMCs and the thyroid. Additionally, <i>IGHV3-23</i> and <i>IGHV4-34</i> may be associated with autoantibody production in GD.https://www.mdpi.com/2073-4409/14/14/1102single-cell RNA sequencingGraves’ diseaseB cellsthyroid autoantigenspeptide stimulationautoantibody production |
| spellingShingle | Genki Kobayashi Takuro Okamura Yoshitaka Hashimoto Kimiko Sakai Madoka Sumi Dan Imai Nobuko Kitagawa Masahide Hamaguchi Michiaki Fukui Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B Cells Cells single-cell RNA sequencing Graves’ disease B cells thyroid autoantigens peptide stimulation autoantibody production |
| title | Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B Cells |
| title_full | Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B Cells |
| title_fullStr | Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B Cells |
| title_full_unstemmed | Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B Cells |
| title_short | Pathogenesis of Graves’ Disease Determined Using Single-Cell Sequencing with Thyroid Autoantigen Peptide Stimulation in B Cells |
| title_sort | pathogenesis of graves disease determined using single cell sequencing with thyroid autoantigen peptide stimulation in b cells |
| topic | single-cell RNA sequencing Graves’ disease B cells thyroid autoantigens peptide stimulation autoantibody production |
| url | https://www.mdpi.com/2073-4409/14/14/1102 |
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