CD4⁺ forkhead box P3 (FOXP3)⁺ regulatory T cells (Tregs) are essential in maintaining immune tolerance and suppressing excessive immune responses. Tregs also contribute to tissue repair processes distinct from their roles in immune suppression. For these reasons, Tregs are candidates for targeted therapies for inflammatory and autoimmune diseases, and in diseases where tissue damage occurs. MT-2 cells, an immortalized Treg-like cell line, offer a model to study Treg biology and their therapeutic potential. In the present study, we use clustered regularly interspaced palindromic repeats (CRISPR)-mediated knockdown of FOXP3 in MT-2 cells to understand the transcriptional and functional changes that occur when FOXP3 is lost and to compare MT-2 cells with primary human Tregs. We demonstrate that loss of FOXP3 affects the transcriptome of MT-2 cells and that FOXP3's potential downstream targets include a wide range of transcripts that participate in the cell cycle, promote growth and contribute to inflammatory processes, but do not wholly simulate previously reported human primary Treg transcriptional changes in the absence of FOXP3. We also demonstrate that FOXP3 regulates cell cycling and proliferation, expression of molecules crucial to Treg function and MT-2 cell–suppressive activities. Thus, MT-2 cells offer opportunities to address regulatory T-cell functions in vitro.

中文翻译：

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存在和不存在叉头盒 P3 (FOXP3) 时 MT-2 Treg 样细胞系的表征

CD4 ⁺叉头框 P3 (FOXP3) ⁺调节性 T 细胞 (Treg) 对于维持免疫耐受和抑制过度免疫反应至关重要。与免疫抑制中的作用不同，Treg 还有助于组织修复过程。由于这些原因，Tregs 是针对炎症和自身免疫性疾病以及发生组织损伤的疾病的靶向治疗的候选者。MT-2 细胞是一种永生化的 Treg 样细胞系，为研究 Treg 生物学及其治疗潜力提供了模型。在本研究中，我们在 MT-2 细胞中使用成簇规则间隔回文重复序列 (CRISPR) 介导的 FOXP3 敲低来了解 FOXP3 丢失时发生的转录和功能变化，并将 MT-2 细胞与原代人类 Tregs 进行比较。我们证明 FOXP3 的缺失会影响 MT-2 细胞的转录组，并且 FOXP3 的潜在下游靶标包括参与细胞周期、促进生长和促进炎症过程的广泛转录本，但并不完全模拟先前报道的人类原发性炎症反应。 FOXP3 缺失时 Treg 转录发生变化。我们还证明 FOXP3 调节细胞周期和增殖、对 Treg 功能和 MT-2 细胞抑制活性至关重要的分子的表达。因此，MT-2 细胞提供了在体外解决调节性 T 细胞功能的机会。