University of Houston, Houston, Texas.
Georgia Institute of Technology, Atlanta.
Arthritis Rheumatol. 2019 Apr;71(4):529-541. doi: 10.1002/art.40772. Epub 2019 Feb 23.
To obtain the comprehensive transcriptome profile of human citrulline-specific B cells from patients with rheumatoid arthritis (RA).
Citrulline- and hemagglutinin-specific B cells were sorted by flow cytometry using peptide-streptavidin conjugates from the peripheral blood of RA patients and healthy individuals. The transcriptome profile of the sorted cells was obtained by RNA-sequencing, and expression of key protein molecules was evaluated by aptamer-based SOMAscan assay and flow cytometry. The ability of these proteins to effect differentiation of osteoclasts and proliferation and migration of synoviocytes was examined by in vitro functional assays.
Citrulline-specific B cells, in comparison to citrulline-negative B cells, from patients with RA differentially expressed the interleukin-15 receptor α (IL-15Rα) gene as well as genes related to protein citrullination and cyclic AMP signaling. In analyses of an independent cohort of cyclic citrullinated peptide-seropositive RA patients, the expression of IL-15Rα protein was enriched in citrulline-specific B cells from the patients' peripheral blood, and surprisingly, all B cells from RA patients were capable of producing the epidermal growth factor ligand amphiregulin (AREG). Production of AREG directly led to increased migration and proliferation of fibroblast-like synoviocytes, and, in combination with anti-citrullinated protein antibodies, led to the increased differentiation of osteoclasts.
To the best of our knowledge, this is the first study to document the whole transcriptome profile of autoreactive B cells in any autoimmune disease. These data identify several genes and pathways that may be targeted by repurposing several US Food and Drug Administration-approved drugs, and could serve as the foundation for the comparative assessment of B cell profiles in other autoimmune diseases.
获取类风湿关节炎（RA）患者中瓜氨酸特异性 B 细胞的综合转录组谱。
使用肽-链霉亲和素缀合物通过流式细胞术从 RA 患者和健康个体的外周血中分选瓜氨酸和血凝集素特异性 B 细胞。通过 RNA 测序获得分选细胞的转录组谱，并通过基于适配体的 SOMAscan 测定法和流式细胞术评估关键蛋白分子的表达。通过体外功能测定法检查这些蛋白对破骨细胞分化以及滑膜细胞增殖和迁移的影响。
与 RA 患者的瓜氨酸阴性 B 细胞相比，瓜氨酸特异性 B 细胞差异表达白细胞介素-15 受体 α（IL-15Rα）基因以及与蛋白瓜氨酸化和环 AMP 信号相关的基因。在对另一组环瓜氨酸肽阳性 RA 患者的分析中，IL-15Rα 蛋白的表达在患者外周血中的瓜氨酸特异性 B 细胞中富集，令人惊讶的是，所有 RA 患者的 B 细胞均能够产生表皮生长因子配体 Amphiregulin（AREG）。AREG 的产生直接导致成纤维样滑膜细胞的迁移和增殖增加，并且与抗瓜氨酸蛋白抗体结合后，导致破骨细胞分化增加。
据我们所知，这是首次在任何自身免疫性疾病中记录自身反应性 B 细胞的全转录组谱的研究。这些数据确定了几个可能通过重新利用几种美国食品和药物管理局批准的药物来靶向的基因和途径，并且可以作为比较评估其他自身免疫性疾病中 B 细胞谱的基础。
