The manufacturing process of Regulatory T (Treg) cells for clinical application begins with the positive selection of CD25+ cells using superparamagnetic iron-oxide nanoparticle (SPION)-conjugated anti-CD25 antibodies (spCD25) and immunomagnetic cell separation technology. Our findings revealed that the interaction of spCD25 with its cell target induced the internalization of the complex spCD25-Interleukin-2 Receptor. Accumulation of intracellular spCD25 triggered oxidative stress, causing delayed Treg expansion and temporary reduction in suppressor activity. This activation delay hindered the efficient generation of clinically competent cells. During this early phase, Treg cells exhibited elevated mitochondrial superoxide and lipid peroxidation levels, with concomitant decrease on mitochondrial respiration rates. The results uncovered the increased mitochondrial unfolded protein response (mitoUPR). This protective, redox-sensitive activity is inherent of Tregs when contrasted with homologous, spCD25-treated, conventional T cells. While the temporary effects of spCD25 on clinically competent cells did not impede their use in a safety/feasibility pilot study with kidney transplant recipients*, it is reasonable to anticipate a potential reduction in their therapeutic efficacy. The mechanistic understanding of the adverse effects triggered by spCD25 is crucial for improving the manufacturing process of clinically competent Treg cells, a pivotal step in the successful implementation of immune cell therapy in transplantation. *Clinical trial registration number NCT03284242 at ClinicalTrials.gov.

中文翻译：

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线粒体氧化应激对调节性 T 细胞制造在移植中临床应用的影响：一项初步研究的结果。


用于临床应用的调节性 T （Treg） 细胞的制造过程从使用超顺磁性氧化铁纳米颗粒 （SPION） 偶联的抗 CD25 抗体 （spCD25） 和免疫磁性细胞分离技术对 CD25+ 细胞进行阳性选择开始。我们的研究结果表明，spCD25 与其细胞靶标的相互作用诱导了复合体 spCD25-白细胞介素-2 受体的内化。细胞内 spCD25 的积累触发了氧化应激，导致 Treg 扩增延迟和抑制因子活性暂时降低。这种激活延迟阻碍了临床感受态细胞的有效生成。在这个早期阶段，Treg 细胞表现出线粒体超氧化物和脂质过氧化水平升高，同时线粒体呼吸速率降低。结果揭示了线粒体未折叠蛋白反应 （mitoUPR） 的增加。与同源的 spCD25 处理的常规 T 细胞相比，这种保护性、氧化还原敏感活性是 Tregs 所固有的。虽然 spCD25 对临床感受态细胞的临时影响并不妨碍它们在肾移植受者的安全性/可行性试点研究*中的使用，但可以合理地预期其治疗效果可能会降低。对 spCD25 引发的不良反应的机制理解对于改进临床感受态 Treg 细胞的制造过程至关重要，这是在移植中成功实施免疫细胞疗法的关键步骤。*临床试验注册号 NCT03284242 ClinicalTrials.gov。