Department of Nanoengineering, University of California, San Diego, La Jolla, CA 92093, USA.
Biomater Sci. 2020 Aug 7;8(15):4186-4198. doi: 10.1039/d0bm00622j. Epub 2020 May 22.
Regulatory T cells (T) are critical mediators of peripheral immune tolerance. T suppress immune activation against self-antigens and are the focus of cell-based therapies for autoimmune diseases. However, T circulate at a very low frequency in blood, limiting the number of cells that can be isolated by leukapheresis. To effectively expand Tex vivo for cell therapy, we report the metabolic modulation of T cells using mono-(6-amino-6-deoxy)-β-cyclodextrin (βCD-NH) encapsulated rapamycin (Rapa). Encapsulating Rapa in β-cyclodextrin increased its aqueous solubility ∼154-fold and maintained bioactivity for at least 30 days. βCD-NH-Rapa complexes (CRCs) enriched the fraction of CD4CD25FoxP3 mouse T (mT) cells and human T (hT) cells up to 6-fold and up to 2-fold respectively and suppressed the overall expansion of effector T cells by 5-fold in both species. Combining CRCs and transforming growth factor beta-1 (TGF-β1) synergistically promoted the expansion of CD4CD25FoxP3 T cells. CRCs significantly reduced the fraction of pro-inflammatory interferon-gamma (IFN-γ) expressing CD4 T cells, suppressing this Th1-associated cytokine while enhancing the fraction of IFN-γ tumor necrosis factor-alpha (TNF-α) expressing CD4 T cells. We developed a model using kinetic rate equations to describe the influence of the initial fraction of naïve T cells on the enrichment of Tin vitro. The model related the differences in the expansion kinetics of mT and hT cells to their susceptibility for immunophenotypic modulation. CRCs may be an effective and potent means for phenotypic modulation of T cells and the enrichment of Tin vitro. Our findings contribute to the development of experimental and analytical techniques for manufacturing T based immunotherapies.
调节性 T 细胞（T）是外周免疫耐受的关键介质。T 抑制针对自身抗原的免疫激活，是自身免疫性疾病细胞治疗的重点。然而，T 在血液中的循环频率非常低，限制了通过白细胞分离术可分离的细胞数量。为了有效地在体外扩增 Tex 用于细胞治疗，我们报告了使用单-（6-氨基-6-去氧）-β-环糊精（βCD-NH）包封雷帕霉素（Rapa）对 T 细胞进行代谢调节。将 Rapa 包封在β-环糊精中可将其水溶解度提高约 154 倍，并至少保持 30 天的生物活性。βCD-NH-Rapa 复合物（CRCs）分别使小鼠 T（mT）和人 T（hT）细胞中 CD4CD25FoxP3 细胞的分数富集了 6 倍和 2 倍，并使两种物种中效应 T 细胞的总体扩增减少了 5 倍。CRCs 与转化生长因子-β1（TGF-β1）联合使用可协同促进 CD4CD25FoxP3 T 细胞的扩增。CRCs 可显著降低表达前炎性干扰素-γ（IFN-γ）的 CD4 T 细胞的分数，抑制这种 Th1 相关细胞因子的表达，同时增强表达 IFN-γ肿瘤坏死因子-α（TNF-α）的 CD4 T 细胞的分数。我们开发了一个使用动力学速率方程的模型来描述初始 naïve T 细胞分数对体外 Tin 富集的影响。该模型将 mT 和 hT 细胞扩增动力学的差异与它们对免疫表型调节的敏感性联系起来。CRCs 可能是一种有效的、有效的方法，用于调节 T 细胞的表型并在体外富集 Tin。我们的研究结果为制造基于 T 的免疫疗法的实验和分析技术的发展做出了贡献。
