The Time Is Crucial for Ex Vivo Expansion of T Regulatory Cells for Therapy
The Time Is Crucial for Ex Vivo Expansion of T Regulatory Cells for Therapy
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DOI：
10.3727/096368911x566217
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发表时间：
2011-01-01
影响因子：
3.3
通讯作者：
Trzonkowski, Piotr
中科院分区：
文献类型：
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作者：
Marek, Natalia;Bieniaszewska, Maria;Trzonkowski, Piotr
体外扩增CD4(+)CD25（高）CD127(-) T调节细胞（Tregs）被认为是人类免疫抑制治疗的有希望的候选者。然而，由于Tregs谱系的可塑性和体外扩增过程中存在的人工环境，Tregs很容易失去抑制活性。在这里，我们跟随扩增CD4（+）、CD25（高）、CD127(-) treg及其初始（CD45RA(+)）和记忆样（CD45RA(-)）亚群，以建立扩增的最佳条件。我们发现，无论表型分类如何，扩增的treg都经历了类似于稳态增殖的变化，并转化为效应记忆样细胞，不仅产生抑制性白细胞介素-10 (IL-10)，还产生IL-6、IL-17和干扰素- γ (ifn - γ)。随着离体时间的延长，在刺激和静止状态下，Tregs都失去了FoxP3的表达和抑制活性。唯一有助于保持Tregs抑制能力的变量是离体培养时间仅为2周。根据我们的研究，CD4(+)CD25（高）CD127(-) treg培养时间不超过2周，可产生最多的高抑制性treg。应进行彻底的质量检查，优先评估FoxP3表达和ifn - γ抑制试验，以评估细胞的抑制活性。
Ex vivo expanded CD4(+)CD25(high)CD127(-) T regulatory cells (Tregs) are recognized as a promising candidate for immunosuppressive therapy in humans. However, due to the plasticity of Tregs lineage and artificial environment present during ex vivo expansion, Tregs easily lose suppressive activity. Here, we followed expanding CD4(+)CD25(high)CD127(-) Tregs and their naive (CD45RA(+)) and memory-like (CD45RA(-)) subsets in order to establish the best conditions of the expansion. We found that, regardless of the phenotype sorted, expanding Tregs were undergoing changes resembling homeostatic proliferation and transformed into effector memory-like cells which produced not only suppressive interleukin-10 (IL-10) but also IL-6, IL-17, and interferon-gamma (IFN-gamma). With the time ex vivo, Tregs were losing the expression of FoxP3 and suppressive activity both when stimulated and when at rest. The only variable that helped preserve suppressive abilities of Tregs was the limitation of the time of ex vivo cultures to 2 weeks only. According to our study, the highest number of highly suppressive Tregs could be yielded with CD4(+)CD25(high)CD127(-) Tregs cultured no longer than 2 weeks. Thorough quality check, preferentially with the assessment of FoxP3 expression and IFN-gamma suppression assay, should be applied to assess suppressive activity of the cells.